Serological selection for cloning of new antigens or genes
E.coli/ phage lysate adsorption serum removes antibodies in serum that immunoreact with bacterial and phage lysin, this reduces the false positive rate, the ser
Topic
Immunological experiments
Techniques for detecting, quantifying and localizing antigens and antibodies — ELISA, Western blotting, immunohistochemistry and flow cytometry. Below are the protocols, FAQs and technical articles in our knowledge base tagged with this topic.
189 resources · protocols, FAQs & articles
Tumor protein immunopreventive assay
Immunological control refers to the application of various types of biological or non-biological biological agents to establish, enhance or inhibit the immune f
Measurement of neutrophil phagocytosis
Measurement of neutrophil phagocytosis can be used to (1) detect phagocytosis in the body and (2) diagnose diseases associated with phagocyte dysfunction.
Functional assay of cytotoxic T cells induced by specific antigens
Specific antigen-induced cytotoxic T cell function assay is mainly applied to (1) the determination of immune cell function (2) the diagnosis of clinical autoim
Enzyme immunoelectrophoresis
Enzyme immunoelectrophoresis can be applied to the analysis and identification of enzyme proteins.
Insulin assay experiment
Radioimmunoassay of insulin can be used for (1) immunolabeling detection techniques using radioisotopes as tracers, and (2) competitive binding of isotope-label
Polyclonal antibody purification assay
The octanoic acid extraction method can be used to isolate and extract Ig from human, rabbit and mouse serum and McAb from hybridoma-induced ascites and culture
Antigen-specific T cell clone preparation assay
The antigen-specific T cell clone preparation assay can be used to: prepare antigen-specific T cell clones.
Cross-protection neutralization experiments
When an animal is infected by a virus, specific neutralizing antibodies are produced in the body and bind specifically to the corresponding virus particles, thu
Radioimmuno-convection electrophoresis assay
When an antibody (or antigen) labeled with a radioactive element is precipitated with the corresponding antigen (or antibody), the precipitation line is confirm
Polyclonal Antibody Preparation Experiment
Antigens are usually composed of multiple antigenic determinants, and the antibody produced by a B-lymphocyte receiving an antigen stimulated by one antigenic d
Estradiol Detection Laboratory
Estradiol (17β-Estradiol) is an important component of natural estrogen, and the detection of estradiol can be used for (1) the determination of hormone additi
Speckled gold immunofiltration assay (DIGFA) assay
The spotted gold immunofiltration assay (DIGFA) is available as a clinical test.
Human Umbilical Vein Endothelial Cell (HUVEC) Culture
Human umbilical vein endothelial cells (HUVEC) can be used (1) as a model cell for in vitro experiments and (2) to study the biological properties of vascular e
ELISPOT
With the wide application of enzyme-linked immunoassay (ELISA) technology in the field of medicine and biology, new breakthroughs have been made in the study of
MTT (tetrazolium salt) colorimetric method
The tetrazolium salt (MTT) colorimetric assay can be applied to (1) detection of cell survival and growth, (2) large-scale screening of antitumor drugs, (3) cyt
Isolation of mouse natural killer cells from mouse spleen
Author: J.E. Collier et al, Translated by Xuedao Cao et al. This experiment is from "Compendium Immunology Laboratory Guide".
Dendritic cell isolation experiments
This unit describes two methods of isolating dendritic cells (DCs): isolating DCs from mouse spleen and obtaining large numbers of DCs from mouse bone marrow pr
Preparation of paraffin sections (hematoxylin-eosin counterstaining method)
Paraffin section making can be used to (1) maintain the normal interrelationships between cells, and can retain the original appearance of cells better and for
Pseudocomplex ciliated columnar epithelium morphologic observation experiments
This experiment is from the website of Basic Medical Experimental Teaching Center of Guiyang College of Traditional Chinese Medicine.
Pregnancy test
Pregnant women's urine to be tested (containing human chorionic gonadotropin, HCG) is interacted with known anti-HCG antibodies, thus inhibiting the binding of
Immunofluorescence complement assay
This experimental method was obtained from the official website of the Fourth Military Medical University
Microimmunoelectrophoresis
Learning the electrophoresis technique of separating proteins in the gel and the immunodiffusion technique of using the specific reaction of antigen and antibod
Antiserum potency assay by two-way immunodiffusion method
The two-way immunodiffusion method can be used to (1) diagnose disease and (2) determine the potency of antibodies.
Experiments on the preparation of immune serum
The preparation of immune serum is a commonly used immunological laboratory technique. Highly efficient and specific immune serum can be used as a reagent for i
B. Lymphocyte membrane surface immunoglobulin examination assay
The test for the examination of immunoglobulins on the membrane surface of B-lymphocytes can be used to: detect the presence of B-lymphocytes.
Phagocytosis assay for phagocytosis
Intraperitoneal injection of sodium thioglycolate in mice can stimulate the aggregation of macrophages. Four days later, mice were injected intraperitoneally wi
Reverse transcription and amplification experiments with low abundance nucleic acids
This method uses only one recombinant enzyme, rTth DNA polymerase, which allows for simultaneous reverse transcription and amplification in a single reaction, t
Immunofluorescence double labeling of tissue sections
Indirect immunofluorescence microscopy allows two or more antigens to be visualized in the same section, at the same time, and can be used for (1) cell labeling
Immunogold labeling of tissue sections
Immunogold labelling technique is a new type of immunolabeling technique applied to antigen-antibody reaction using colloidal gold as tracer. Colloidal gold is
Immunofluorescence labeling assay of streptavidin-biotin conjugates
Immunofluorescent labeling is a commonly used immunological experimental method in microbiology, immunology, pathology and immunohistochemistry.
Immunofluorescence labeling of tissue sections
The immunofluorescence technique is the labeling of a known antibody or antigen molecule with fluorescein, which can be used for (1) examination of antigen-anti
Immunofluorescence labeling of suspension cells
Immunofluorescence technique is to label a known antibody or antigen molecule with fluorescein, and when it reacts with its corresponding antigen or antibody, a
Immunofluorescence labeling assay of monolayer growth cells
Immunofluorescent labeling is a commonly used immunological experimental method in microbiology, immunology, pathology and immunohistochemistry.
Frozen tissue fixation and sucrose immersion experiments
Tissue morphology is usually good for frozen sections if the tissue is fixed and macerated with sucrose prior to sectioning. Source: Practical Laboratory Techni
Mixed lymphocyte culture assay
Mixed lymphocyte culture (MLC) or mixed lymphocyte reaction (MLR) was previously used for tissue matching prior to organ transplantation to determine the degree
Assay of killer cell function
Killing function is an important aspect of the body's immune function, and there are a variety of target cells with killing function in the immune system, such
Lymphocyte transformation assay
Lymphocyte transformation assays are used in clinical practice to (1) reflect the response function of the overall level of T-cells in the subject, to determine
Leukocyte movement inhibition assay
Sensitized T-lymphocytes react with the corresponding antigen, causing metabolic activation and the release of a variety of biologically active substances. Amon
Detection of antibody-producing cells-Hemolytic vacuole assay
Spleens of mice immunized with sheep cells were taken and spleen cell suspension was made, which was combined with a certain amount of indicator cells (sheep ce
complement binding reaction assay
When an antigen binds to its corresponding antibody, the resulting antigen-antibody complex is able to absorb the complement from the solution, which is called
hemolytic reaction assay
Immune serum meets with its corresponding antigenic cells, blood cells, bacteria and their tissue cells, and a cytolytic reaction can occur with the participati
agglutination assay
Many of the Enterobacteriaceae have partially identical antigenic structures. Because of this one bacterium can cross agglutinate with the corresponding antiser
Rocket electrophoresis experiment
A fixed amount of antigen is added to the agarose gel containing a certain amount of the corresponding antibody, under the action of the electric field, causing
Convection immunoelectrophoresis assay
Convection immunoelectrophoresis is essentially an immunodiffusion technique with directional acceleration, the basic principle of which is: two rows of holes a
Observation experiments on immune-related cell morphology
All living things are made up of cells, only the size and shape of the cells vary from organism to organism. Some cells are visible to the human eye, such as th
Technical route and basic fabrication process of tissue microarray preparation
Tissue microarrays are made by punching holes with a fine needle in a tissue microarray machine, collecting tens to hundreds of small cylindrical tissues (tissu
Serological screening for cloning of new antigens/new genes
Screening for new antigens and genes through a system of studies of the body's immune response and metabolic reactions centered on various reactions to serum.
Immunization program for laboratory animals
The basic theories and techniques of immunology are used to diagnose, prevent and treat diseases, and on this basis, artificial methods are used to input into t
Identification and characterization experiments of human tumor antigens
Human tumor antigens can be (1) used for tumor diagnosis and (2) immunotherapy.
immunohistochemical staining
Immunohistochemical staining refers to the combination of fluorescent or colorable chemicals on antibodies to detect the presence of target antigens in cells or
Complement assay experiment
Complement assay is the measurement of total complement hemolytic activity and is used to detect various acute inflammatory conditions.
Indirect agglutination inhibition test
After mixing the sample to be tested with the known antibody for a certain period of time, and then mixing it with the latex particles sensitized by the corresp
Immunological organ recognition assay
The organs of immunity include the central and peripheral organs of immunity. In mammals, the central immune organs include the thymus and bone marrow; in birds
Magnetic resonance imaging diagnosis of chordoma and differential diagnostic tests
Chordoma arises from residual embryonic chord or ectopic chord and occurs only in the median bones, preferably at the ends of the spine. Pathologically, there a
Hemolytic vacuole formation test
The hemolytic vacuole formation test is an in vitro method for the detection of individual antibody-forming cells (plasma cells), and is therefore also known as
Enzyme-linked immunosorbent assay (ELISA) for the determination of antibodies against S. typhi "O"
Enzyme-linked immunosorbent assay (ELISA) is a method of detecting unknown antigens or antibodies with enzyme-labeled antibodies (or SPA). This method is highly
Tumor protein vaccine prophylactic trials
Tumor protein vaccine prophylactic trials are designed to examine whether the vaccine's targets are directed against a wide range of antigens, can cover 90% or
one-way immunodiffusion assay
A single immunodiffusion assay can be used to: measure IgG, IgA and IgM levels in the serum of normal people or patients.
nucleic acid immunoassay
The ways to enhance the immune response with antigen-expressing DNA are described. Two methods of nucleic acid immunization are described: (i) injection of sali
Isolation experiments of single nucleated cells in mice
The basic techniques for preparing cell suspensions from mouse lymphoid organs are first described, including the removal of erythrocytes and dead cells The met
Determination of guinea pig T-lymphocytes -Rabbit erythrocyte rosette assay
Human T and B lymphocytes have different receptors on their surface. Since human T lymphocytes have receptors on their surface that bind to sheep erythrocytes a
Immunoperoxidase labeling assay of tissue sections
This is a very sensitive method. The antibody used to form the HRPO-antiperoxidase (PAP) complex must be of the same type as the primary antibody recognized by
Separation of T cells and B cells by magnetic bead method
This unit describes the separation of T cells, B cells, and T cell subpopulations by magnetic separation techniques. Author: J.E. Collier et al, Translator: Xui
Cloning, Expression and Modification of Antibody Variable Regions (V Regions)
PCR-based cloning schemes utilize conserved sequences in the framework and pilot regions of the variable region, and although primers can be synthesized with co
Hydrolysis of immunoglobulin G (IgG) fragments
When a new antibody needs to be fragmented, it is often necessary to perform a touch-and-go test. When choosing the reaction time, it is preferable to keep part
Human α2a, α2b Interferon Assay
An indispensable part of the production process of recombinant human α2a/α2b interferon is the quality control of the product, and the bioactivity o
Soluble Interleukin 2 Receptor Assay
The level of concentration of soluble interleukin 2 receptor SIL-2R is closely related to many diseases, such as adult T-lymphoblastic leukemia AIDS, B-cell slo
IL-10 biological activity assay
In the presence of mouse IL-4 (Mil-4), IL-10 stimulated proliferation of the D36 mouse mast cell line. In contrast, single mIL4 or hIL-10 had only very low prol
IL-6 biological activity assay
MH60.BSF2 is an IL-6-dependent cell line, and the amount of MH60.BSF2 cell proliferation was positively compared with the biological activity of IL-6.
IL-2 biological activity assay
Interleukin-2 (IL-2) is a cell proliferation factor secreted by activated helper T cells, which has the effect of promoting T cell proliferation and maintaining
T and B lymphocyte separation assay
Lymphocytes are divided into two main subpopulations, T and B lymphocytes, which have different properties and functions and are used for (1) immunological stud
Immunocytochemistry ICC
Localized detection of antigen expression
electrochemiluminescence immunoassay (ECLI)
labeled immunoassay
Experiments on the electron microscopic display of cell membrane glycans
Cell membrane glycans electron microscopy display experiments can be used to: observe glycoproteins and glycolipids on cell membranes. The more commonly used di
Acetylcholine receptor display cytochemistry experiments
Acetylcholine receptors are distributed at neuromuscular junctions, etc. Snake toxin binds tightly to them and connects to peroxidase, and the distribution of a
Tracer cytochemistry experiments
Since high electron density tracers can easily diffuse in the interstitial space of cells, and tracers can also enter into cells if cell damage occurs, this met
Ion cytochemistry experiment
Ion cytochemistry can be used to show the distribution of intracellular ion localization. Currently, it is more commonly used to show the distribution of intrac
Immunoelectron microscopy specimen fixation experiment
Immunoelectron microscopy specimens need to be fixed after sampling. Source of content Immunohistochemistry Laboratory Techniques and Applications
Electron microscopy enzyme cytochemistry experiment
Currently there are three main classes of enzymes that can be localized by electron microscopy; they are hydrolytic enzymes, oxidative enzymes, and transferase
mast cell staining assay
Mast cells are derived from undifferentiated mesenchymal cells and are found in tissues such as perivascular submucosa, loose connective tissues, tumor tissues,
Perfusion fixation experiments
Source: Practical Experimental Techniques in Neurobiology, Fourth Military Medical University Press
IHC sensitization test
There are several ways to increase the sensitivity of IHC. One is to rely on various IHC pre-treatments, such as enzyme digestion and antigen repair at high tem
two-way immunodiffusion assay (BIDA)
The diffusion of antigen and antibody in a gel and the precipitation reaction is called immunodiffusion. An antigen and its corresponding antibody are placed in
Experiments on the preparation of antigens and immune sera
Injecting the antigen into the animal body can stimulate the animal's B lymphocytes to transform into plasma cells and produce specific antibodies. When a large
Immune molecules and receptor engineering
A method for the preparation of soluble M H C -I g dimers is described, and a method for the preparation of multimeric M H C -I/peptide complexes is presented.
Isolation of human intestinal mucosal single nucleated cells
Isolation of single nucleated cells from human intestinal mucosa Author: J.E. Colligan et al, Translated by Xuitao Cao et al. This experiment is from the "Compe
Cloning and Expansion of Human T Cells
Induction of antigen-specific T cells Author: J.E. Colligan et al, Translated by Xuitao Cao et al. This experiment is from "Comprehensive Immunology Laboratory
Detection of T-cell killing function
The assay of T-cell killing function can be applied to (1) immune cell function determination (2) clinical immunodiagnosis.
Isolation and purification of T cell subsets
Isolated T cells are categorized into CD4+ and CD8+ T cells using anti-CD8 antibody. Since both types of cells can be recovered, this method is superior to anti
Isolation of single nucleated cells from peripheral and umbilical cord blood
Peripheral blood mononuclear cells can be used in: (1) immunology (including autoimmune diseases), infectious diseases, hematologic malignancies, vaccine develo
Selection and preparation of antigen-presenting cells
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Activation of mouse macrophages
Macrophage activation goes through two phases: a sensitization phase and an activation phase. While most protocols for macrophage activation use a one-step appr
Detection of intracellular factors by flow cytometry
This unit describes the use of 4-color fluorescence to analyze the levels of IFN-7, IL-4 and IL-2 expressed in CD4+ T cells. (can be used to calculate T h l and
Immunofluorescence and cell separation
Flow cytometry is used to analyze membrane surface antigens and intracellular antigens expressed by cells, and is capable of specifically distinguishing differe
Separation and application of lipid rafts
Lipid rafts are important in immunology because T-cell receptors and several other important T-cell signaling proteins are present in lipid rafts or are recruit
Preparation of antibodies recognizing specific tyrosine phosphorylated polypeptides
Antibodies against phosphopeptides recognize proteins that are only in the phosphorylated state. The antibody does not cross-react with unphosphorylated protein
Detection of apoptosis and other forms of cell death
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
T lymphocyte proliferation assay
This module introduces the mouse T lymphocyte proliferation assay. Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from the "Comp
Cytochemical experiments on lectin-displaying glycans
Lectin is a non-immunogenic glycoprotein that binds specifically to sugars, e.g., lectin from kidney beans binds only to glucose and mannose, and lectin from wh
IL-4 biological activity assay
IL-4 is mainly a cytokine produced by the activation of TH cells.CT.4S cells, an IL-4-dependent cell line, show hyporesponsiveness to IL-2, and their proliferat
IL-8 Biological Activity Assay
There are activating and chemotactic effects on neutrophils in IL-8, and the biological activity of IL-8, i.e., chemotactic activity on neutrophils, can be dete
Experiments on cyclic precipitation reactions
The cyclic precipitation reaction is widely used in forensic medicine for the identification of bloodstains and the determination of food adulteration. Usually
B Assays of cellular function
Author: J.E. Collier et al, Translated by Xuedao Cao et al. This experiment is from "Compendium Immunology Laboratory Guide".
Target cell killing assay: Cr51 method
Detection of killing activity of effector cells
Calcium staining assay
Calcium is deposited in bone tissue or pathologically altered tissue, most often in calcified tissue such as tissue necrosis, atherosclerosis and tumors. In the
B Early events in lymphocyte activation
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Phenotyping of Human Monocytes/Macrophages
To provide a phenotypic assay for human monocytes/macrophages. Author: J.E. Colligan et al, Translator: Xuitao Cao et al. This experiment is from the "Comprehen
Interleukin 8 (IL- 8) Assay
Il-8 is a polypeptide with a molecular weight of 8-10 kD, which has chemotactic effects on neutrophils, T lymphocytes, and alkaliphilic granulocytes, and activa
IL-1 biological activity assay
Interleukin-1 (IL-1) is a cytokine mainly synthesized and secreted by activated monocyte-macrophage cells, and has various biological functions such as activati
Preparation of polyclonal antibodies against phosphopeptides.
It is possible to prepare anti-phosphopeptide antibodies (i.e., antibodies that recognize phosphorylated peptides) that recognize only proteins in the phosphory
In situ PCR amplification of DNA and RNA target sequences
In situ PCR, a technique established by Hasse in 1990, involves performing PCR reactions in tissue cells. It combines the advantages of in situ hybridization wi
Monoclonal antibody ascites preparation experiment
The primary advantage of using monoclonal antibodies (MAb) over polyclonal antisera is the possibility of obtaining large quantities of specific monoclonal anti
B Phenotypic detection of cells
There are many membrane molecules on the surface of B cells, including cell surface differentiation antigen and model immunoglobulin (smlg). At different stages
Isolation and purification of human metaphase stromal cells
Endometrial metamorphosis is a critical step for embryo implantation and successful pregnancy. The cellular composition of the metaphase tissue is quite complex
Isolation and culture of human chorionic extrachorionic trophoblast cells
The normal exercise of trophoblast biological functions is a key component in maintaining a normal pregnancy. After embryo implantation, trophoblast cells divid
Cultured dendritic cells from mouse bone marrow precursor cells
Dendritic cells (DCs) are very few in number in vivo (less than 1% of splenic adherent cells), and the time-consuming isolation of DCs from vivo and their low c
Isolation and purification of mouse peritoneal macrophages
Macrophages (MΦ) are mature cells formed by the migration of blood monocytes into tissues, and are widely distributed in the spleen, liver, abdominal cavity, n
B Cell activation
B cells are activated and proliferate upon activation by specific antigens, anti-Ig antibodies, and/or mitogens.
Experimental effects of anticancer drugs on ⅡDNA topoisomerase
The effect of anticancer drugs on DNA topoisomerase activity assay was performed using agarose gel electrophoresis to determine the effect of anticancer drugs o
Isolation and purification of IgG
IgG isolation and purification experiments can be used to: prepare enzyme labeled antibodies or fluorescent antibodies.
Preparation of EBV-transformed B cells
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of NK/LAK cell killing activity
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
In vivo induction and detection of antibodies
Detection of Antibody Production in Vivo by ELISA by J.E. Colligan et al, Translated by Xuitao Cao et al. This experiment is from the "Compendium of Immunology
ELISA for detection of antibodies
This method detects IgG, I g M and IgA in serum or cell culture supernatants (Fig. 8. 1 0 . 1 ). The color of the liquid in the assay wells is directly proporti
B Detection of cell-secreted antibodies
The following is an example of mitogen-induced antibody production by B cells using American continental protein. This method has been adapted for use with a va
Detection of human lymphocytes in whole blood by flow cytometry
This method analyzes whole blood cells compared to analyzing Ficoll-isolated single nucleated cells, thus taking less time, requiring a smaller sample size, and
Isolation of B-cell subpopulations by immunomagnetic bead assay
Anti-CD 19 labeled B cells, which have relatively low signal intensity, show mitogenic and antibody secretory responses similar to those obtained by negative sc
Presentation of exogenous antigens to T cells
For granular antigens or liposome-encapsulated antigens, macrophages were applied as antigen-presenting cells. Author: J.E. Colligan et al, Translator: Xuitao C
Detection of Fcγ Receptor-Mediated Adhesion and Phagocytosis
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of anti-tumor activity of macrophages
Mouse macrophage cell lines with tumor cell killing activity, such as A N A -1 and G G 2E E . Mouse primary macrophages can be freshly isolated. Author: J.E. Co
Detection of phagocytosis and killing function of macrophages
An assay for macrophage phagocytosis and killing function. Author: J.E. Colligan et al, Translator: Xuitao Cao et al. This experiment is from "Compendium Immuno
Detection of Nitric Oxide in Macrophages
This protocol reflects the level of nitric oxide secretion by detecting nitrite in the culture supernatants of activated macrophages (walled or non-walled) Auth
Phenotypic assay of mouse macrophages
Due to the lack of characteristic surface markers of macrophages or macrophages at different stages of differentiation, macrophages are currently identified bas
Isolation of mouse macrophages
Introduction of a method for the isolation of mouse macrophages by J.E. Colligan et al, translated by Xuitao Cao et al. This experiment is from the "Compendium
ELISPOT method for detection of cytokine-secreting cells
Enzyme-linked immunospot test (ELISPOT) is based on the principle that cells that secrete certain protein molecules (e.g. cytokines) are able to secrete a relat
Detection of cytokine secretion and isolation of cytokine-secreting cells by flow cytometry
This protocol is for the detection of cytokine production by activated antigen-specific T cells in vitro. If other stimulants or stimulated cells are used in vi
Gamma interferon assay
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of IL-15
The ELISA method can be used to detect human IL-15 in culture supernatants and serum samples. This method can also be used for the detection of monkey IL-15, bu
Detection of IL-13
ELISA is highly specific for human or mouse IL-13 and does not recognize other cytokines in the sample. Author: J.E. Colligan et al, Translator: Xuitao Cao et a
IL-12 detection assay
This is a method for the specific detection of IL-12 heterodimers. Since IL-12 p4 0 monomer cannot be detected, this method is able to detect IL-12 p7 5 heterod
IL-2 and IL-4 assays
A method is described for the detection and quantification of one of the cytokines in the presence of the other using neutralizing antibodies to murine IL-2 and
Analysis of mitogen-activated protein kinase (MAPK) activity in T cells
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of tyrosine protein kinase by immunocomplex methods
The low abundance of tyrosine protein kinase (T P K ) expression and its rapid activation and inactivation make it difficult to purify using conventional bioche
Preparation of mouse T-cell hybrid tumors
T-cell hybridomas can be obtained by fusing activated T cells with tumor cells. Heterogeneous hybridomas can be cloned by limited dilution to obtain hybridomas
Establishment of T-cell clones
This unit provides a method for establishing specific T cell clones. Note that the key to the success of this method is the selection and preparation of conditi
BrdU assays for d-cell and B-cell proliferation
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Natural killer cell clearance in mice
NK cells are usually injected intravenously; in vivo clearance can be achieved by intraperitoneal injection of antibodies against NK cells, such as PK136 monocl
Induction and detection of immunoglobulin class switching
During the immune response, B cells stimulated by different antigens can switch to express different heavy chain constant region genes and change the type of im
Macrophage chemotaxis assay
Macrophage migration is the primary condition for its immune activity at the site of infection and injury, and macrophage chemotaxis reflects its migration abil
Blotting assay for anti-phosphorylated tyrosine
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of IL-18
Newly synthesized IL-18 is a precursor molecule to the signal peptide, which is enzymatically cleaved in order to activate it. In this ELISA method, in order to
Induction and detection of cytotoxic T lymphocytes
This unit describes several experimental protocols for inducing cytotoxic T lymphocytes and assaying their activity Remember that, in principle, any kind of cel
B Identification of functional subpopulations of cells
Peripheral mature B cells can be divided into several subpopulations. Traditionally, B cells have been divided into two subpopulations, CD5+ B1 cells and CD5 B2
Induction and characterization of macrophage polarization in mice
Macrophages can undergo functional activation (polarization) of different nature into M1 and M2 subpopulations with different surface molecular and functional c
Detection of lymphocyte migration and proliferation using the intracellular fluorescent dye CFSE
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
IL-10 detection assay
This protocol uses an antibody specific for mouse IL- 1 0 in the E LISA method to detect mouse IL- 1 0 , and does not detect human or viral IL-10. IL-10 from ot
Analysis of inositol phospholipid turnover during lymphocyte activation
Receptor-mediated activation of phospholipase C (P L C) induces hydrolysis of membrane inositol phospholipids to produce diacylglycerol (D A G) and water-solubl
Isolation and expansion of human NK T cells
This protocol allows the identification and quantitative analysis of trace amounts of N K T cells. Effective measures should be taken to block non-specific stai
Detection of antinuclear antibodies in serum by immunofluorescence technique assay
Antinuclear antibody (ANA) is an autoantibody directed against the nuclear components of cells, with no organ or germline specificity. A positive ANA is indicat
B Proliferation assays for cell function
Author: J.E. Colligan et al, Translator: Xuitao Cao et al, This experiment is from "Compendium Immunology Laboratory Guide".
Detection of α-, β- and γ-interferon-induced antiviral activity
The cells used in the assay are mouse L 929 fibroblasts, which have a high sensitivity to interferon and V S V . The cells should be derived from laboratories t
immunoprecipitation mass spectrometry
Immunoprecipitation is a technique used to enrich or purify specific proteins from complex mixtures, which can then be combined with mass spectrometry to identi
Cell cycle analysis of desynchronized cell populations
Information on cell cycle dynamics can be obtained by labeling 5' BrdUrd, which can be incorporated into the thymidine site of DNA. Detection of BrdUrd in DNA a
Mammalian protein-protein interaction traps based on fluorescence-activated cell sorting technology
The Mammalian Protein-Protein Interaction Trap System (MAPPIT) is a two-hybrid system designed based on the understanding of the signaling mechanism of type I c
Combined application of flow cytometry and gene chip methods to detect transcriptional profiles
The two technologies, flow cytometry and gene expression profiling, share many common features: they involve complex, highly sophisticated equipment; they can p
Characterization of hematopoietic stem cells
Hoechst 33342 and Rhodamine123 Staining for Hematopoietic Stem Cell Characterization Given that the in vitro in vivo dyes Rhodamine123 (Rh123) and Hoechst 33342
Multi-parameter analysis of apoptosis using flow and image cytometry
Apoptosis regulates the dynamic balance of cells, and the multiparametric nature of flow cytometry allows for the simultaneous determination of several apoptoti
Evaluation of lymphocyte-mediated cytotoxicity using flow cytometry
Cytotoxicity of lymphocytes, is the killing of target cells by the release of particles containing perforin and granzyme and/or through the interaction of Fas-F
Multi-parameter data acquisition and analysis experiments on leukocytes
Several software packages are now available for flow cytometry data analysis. Although each one has a different approach in processing the data, the basic strat
Preparation of monoclonal antibodies
Hybridoma cells are formed by fusing an immune animal B lymphocyte that produces a single antibody with myeloma cells by cell fusion techniques (e.g., PEG-induc
Isolation of peripheral blood single nucleated cells
Immune cell isolation techniques are developing in the direction of more and more "fine", immune cell subpopulation and even micro-immune cell isolation and ide
Induction and isolation of human blood mononuclear macrophages
Macrophages (MΦ) are mature cells formed by the migration of blood monocytes into tissues, and are widely distributed in the spleen, liver, abdominal cavity, n
CFSE assay for human peripheral blood T cell expansion in vitro
CFSE detection of human peripheral blood T cell expansion in vitro refers to the use of CFSE to fluorescently label live cells, which can be used for the detect
Antibody potency assay
Antibody potency assay is used to test the potency of immune serum. Currently there are three main methods used to determine antibody potency: tube agglutinatio
Chemiluminescence immunoassay experiment
Chemiluminescence (CL) refers to the phenomenon of light radiation that accompanies the process of a chemical reaction. Most chemiluminescence is the luminescen
Flow cytometric analysis of fluorescence resonance energy transfer
The study of various cellular life processes has been made easier by the detection of intermolecular interactions in vivo and in vitro through a variety of meth
Sorting of live, infectious cells by BSL-3 laboratory
As the need for high-speed sorting of viable, infectious and therapeutic cell samples increases, so does the importance of protecting the flow cytometer operato
Flow cytometry analysis of cytokines
Cytokineflow cytometry (CFC) is the collective term for flow cytometry that applies anti-cytokine antibodies to analyze cytokines that are markers of cell activ
Analysis of DNA content in solid tumors
Flow cytometry DNA content assays provide a rapid and credible method of analyzing DNA ploidy and cell proliferation scores in tumor specimens. Both preserved a
Simultaneous flow analysis of DNA and RNA
Simultaneous DNA and RNA analysis by flow cytometry provides information on the DNA content of the cell and the transcriptional status of the cell. This assay d
Isolation and purification of human metaphase immune cells
The metaphase immune cell population is the biological basis of maternal-fetal immune tolerance, and its composition is extremely special, mainly composed of sp
Measurement of immune cell phagocytosis
The body has phagocytosis (phagocytosis) function of the cells called phagocytes, including monocyte macrophages and neutrophils two categories. Phagocytes phag
In vivo activation of macrophages
Macrophages are important natural immune cells in vivo, and in vitro detection of their function does not fully reflect their in vivo behavior, while detection
Detection and enrichment of hematopoietic stem cells by marginal group cell phenotyping
Detection and enrichment of hematopoietic stem cells by marginal group cell phenotyping is based on the isolation of hematopoietic stem cells from mouse bone ma
Monoclonal antibody identification test
The identification of monoclonal antibodies is similar to that of polyclonal antibodies, and the methods used are common; however, the identification of some of
Frequency of antigen-specific T-cell precursor proliferation detected using cellular tracer dyes
T cells recognize specific antigenic molecules (epitopes) through T cell antigen recognition receptors, and although the body's T cell pool can respond to a var
Experimental phenotypic and functional analysis of CD34NEG hematopoietic precursor cells mobilized from peripheral hematopoietic pools
Phenotypic and functional analysis of CD34NEG hematopoietic precursor cells mobilized from peripheral hematopoietic pools is an experiment to study the modulati
Spot immunofiltration assay
A detection test that utilizes antigens or antibodies to carry out a corresponding specific immune response, and is one of the main methods used in immediate te
Related topics
Techniques often explored alongside immunological experiments.