In vitro T-cell activation commonly uses CD3/CD28 antibodies or PMA/Ionomycin stimulation to analyze TCR signaling, cell proliferation, metabolic reprogramming, cytokine secretion, and effector differentiation.
Flow Cytometry
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.
PBS Only antibodies refer to antibody preparations formulated solely in PBS as the storage buffer, typically without the addition of stabilizers or preservatives such as BSA, gelatin, glycerol, or sodium azide. Compared with conventional antibody preparations containing protein stabilizers or ...
Once a cell sample enters a preservation system, the analytical result is no longer determined solely by its original biological state, but is instead jointly influenced by the anticoagulation mode, osmotic environment, buffering conditions, degree of metabolic suppression, and processing ...
Fluorophores are light-emitting molecules or materials that absorb energy upon excitation at specific wavelengths and emit photons at longer wavelengths. They are widely used in cell imaging, flow cytometry, immunoassays, nucleic-acid quantification, protein–protein interaction analysis, and ...
Experimental Steps for Methanol Permeabilization & Triton X-100 Permeabilization
The detection of cell surface proteins mainly targets antigens on the cell membrane. Antibodies can directly bind to antigens on the cell surface, so fixation and permeabilization treatment are usually not required. The experimental operation is relatively simple, and the main focus is on the ...
Flow cytometry is a powerful technique that is often used to analyze surface and internal antigens of cells. When detecting intracellular antigens, cells usually need to be fixed and permeabilized.
An isotype control antibody appears like a typical antibody but it does not attach itself to a specific target protein like the primary antibody used in the experiment.
Flow cytometry (FCM) is a single-cell quantitative analysis and sorting technique using flow cytometry. Flow cytometry is a product of highly developed and comprehensive use of monoclonal antibodies and immunocytochemistry, lasers and computer science, etc. It can effectively differentiate ...
Common Problems and Solutions for Flow Experiments
Flow cytometric surface staining procedure
Intranuclear Factor Staining Procedure
Preparation and precautions for human peripheral blood PBMCs
Procedures and precautions for the preparation of human peripheral blood monocyte suspensions
Preparation of mouse ascites and single cell suspension and precautions
Mouse Bone Marrow Single Cell Suspension Preparation Procedures and Precautions
Preparation and precautions for mouse lymph node single-cell suspensions
Procedures and precautions for the preparation of mouse peripheral blood single-cell suspensions
Procedures and precautions for the preparation of mouse spleen single-cell suspensions
Procedures and precautions for preparation of mouse thymus single cell suspension
Preparation and precautions for single-cell suspensions of mouse tumor samples
Single cells are a prerequisite for the analysis and detection of cells by flow cytometry. In the application of flow cytometry, the preparation of qualified dispersed single cells is an important part of the flow cytometry sample preparation technique. It requires both the dispersion of tissues ...
Cell nuclear factor staining process
Characterization and application scenarios of various enzymes in the preparation of tissue single-cell suspensions
Preservation methods and precautions for common flow-through samples
Common sense in the purchase of flow antibodies
Selection of fluorescent dyes for multicolor flow-through experiments
How do you really choose flow antibodies? What to look for?
Selection of Mouse-Specific Cell Surface Markers
First, we should harvest our cells or tissue and prepare a single-cell suspension. We will then transfer our single-cell suspension into 96-well plates, test tubes, or polystyrene round bottom tubes, depending on the number of cells and volumes being used.
Flow cytometry enables the separation of a suspended sample of cells or particles through a thin, rapidly moving liquid stream. As the sample passes through a laser, it can detect the size, granularity, and fluorescent properties of each individual cell or particle. This technique allows for the ...
Techniques often explored alongside immunological experiments.
