I-A/I-E Rat mAb - Primary antibody, specific to I-A/I-E, Rat IgG2b

Cat. No.: Ab115281

GRADE & PURITY Carrier Free, Validated, ExactAb™, Azide Free, PBS Only, ≥95%(SDS-PAGE), See COA

Application

Animal Model, Blocking, Flow Cytometry, Functional Assay, IF/ICC, IP, WB

Host species

Rat

Reactive species

Mouse

Isotype

Rat IgG2b

Clone Number

M5/114

Conjugation

Unconjugated

Overview

Rat anti Mouse I-A/I-E Antibody, Monoclonal (M5/114), could be used for in vivo MHC II blockade, Functional assays, ICC, IF, WB, IP, Flow and so on.

Application:
in vivo MHC II blockade: Use at an assay dependent concentration. 

Functional assays: Use at an assay dependent concentration. 

ICC/IF: Use at an assay dependent concentration. 

WB: Use at an assay dependent concentration. 

IP: Use at an assay dependent concentration. 

Flow: Use at an assay dependent concentration.

Protein Function
Binds peptides derived from antigens that access the endocytic route of antigen presenting cells (APC) and presents them on the cell surface for recognition by the CD4 T-cells. The peptide binding cleft accomodates peptides of 10-30 residues. The peptides presented by MHC class II molecules are generated mostly by degradation of proteins that access the endocytic route, where they are processed by lysosomal proteases and other hydrolases. Exogenous antigens that have been endocytosed by the APC are thus readily available for presentation via MHC II molecules, and for this reason this antigen presentation pathway is usually referred to as exogenous. As membrane proteins on their way to degradation in lysosomes as part of their normal turn-over are also contained in the endosomal/lysosomal compartments, exogenous antigens must compete with those derived from endogenous components. Autophagy is also a source of endogenous peptides, autophagosomes constitutively fuse with MHC class II loading compartments. In addition to APCs, other cells of the gastrointestinal tract, such as epithelial cells, express MHC class II molecules and CD74 and act as APCs, which is an unusual trait of the GI tract. To produce a MHC class II molecule that presents an antigen, three MHC class II molecules (heterodimers of an alpha and a beta chain) associate with a CD74 trimer in the ER to form an heterononamer. Soon after the entry of this complex into the endosomal/lysosomal system where antigen processing occurs, CD74 undergoes a sequential degradation by various proteases, including CTSS and CTSL, leaving a small fragment termed CLIP (class-II-associated invariant chain peptide). The removal of CLIP is facilitated by HLA-DM via direct binding to the alpha-beta-CLIP complex so that CLIP is released. HLA-DM stabilizes MHC class II molecules until primary high affinity antigenic peptides are bound. The MHC II molecule bound to a peptide is then transported to the cell membrane surface. In B cells, the interaction between HLA-DM and MHC class II molecules is regulated by HLA-DO. Primary dendritic cells (DCs) also to express HLA-DO. Lysosomal miroenvironment has been implicated in the regulation of antigen loading into MHC II molecules, increased acidification produces increased proteolysis and efficient peptide loading.

Specifications

Product Name

I-A/I-E Rat mAb - Primary antibody, specific to I-A/I-E, Rat IgG2b

Synonyms

IAb | Ia2 | Ia-2 | Abeta | H-2Ab | H2-Ab | Rmcs1 | I-Abeta | Eb | Ia4 | H2Eb | Ia-4 | H-2Eb | E-alpha-f | H-2 class II histocompatibility antigen, E-K alpha chain | H-2 class II histocompatibility antigen, E-U alpha chain | H-2E | H2-Ea | H-2Ea | H2-Ea-ps

Specifications & Purity

Carrier Free, ExactAb™, Azide Free, Validated, PBS Only, ≥95%(SDS-PAGE), See COA

Host species

Rat

Specificity

I-A/I-E

Immunogen

Activated C57BL/6 mouse spleen cells

Conjugation

Unconjugated

Grade

Azide Free, Carrier Free, ExactAb™, PBS Only, Validated

Isotype

Rat IgG2b

Light Chain Type

kappa

SDS-PAGE

150 kDa

Purification Method

Protein G purified

Concentration

See COA

Storage and Shipping

Shape

Liquid

Storage

Store at 2-8°C,Do not freeze

Shipped In

Wet ice,Do not freeze

Stability And Storage

Shipped at 4°C. Store at 4°C long term (12 months). Do not freeze. Upon receipt, it is recommended to aliquot.

Images

Flow Cytometry: I-A/I-E Rat mAb (Ab115281)

I-A/I-E Rat mAb (Ab115281) - Flow Cytometry
Flow Cytometry analysis of mouse splenocytes cells labelling I-A/I-E with I-A/I-E Rat mAb (Ab115281) conjugated with biotin (right panel) at 0.5 μg/mL for 1 hour at 4°C, compared with Rat IgG Isotype Control Antibody (Biotin) (Ab180310) (left panel) and detected with SA-PE (rp156150). 

Flow Cytometry: I-A/I-E Rat mAb (Ab115281)

I-A/I-E Rat mAb (Ab115281) - Flow Cytometry
Flow Cytometry analysis of mouse splenocytes cells labelling I-A/I-E (red) with I-A/I-E Rat mAb (Ab115281) conjugated with biotin at 0.5 μg/mL for 1 hour at 4°C, and detected with SA-PE (rp156150). Blue - Rat IgG Isotype Control Antibody (Biotin) (Ab180310). Black - Unlabelled control, cells without incubation with primary antibody.

Application

• Application: Dilution info
• WB:

  Use at an assay dependent concentration.;

• Functional Assay:

  Use at an assay dependent concentration.;

• IP:

  Use at an assay dependent concentration.

• IF/ICC:

  Use at an assay dependent concentration.;

• Flow Cytometry:

  Use at an assay dependent concentration.

• in vivo MHC II blockade:

  Use at an assay dependent concentration;