Overview

  • Product name

    Anti-MHC Class II antibody [M5/114]
    See all MHC Class II primary antibodies
  • Description

    Rat monoclonal [M5/114] to MHC Class II
  • Host species

    Rat
  • Specificity

    ab139365 reacts with murine MHC class II glycoproteins. It recognizes a shared determinant on I-Ab, I-Ad, I-Aq, and I-Ed, I-Ek alloantigens, but it does not react with I-Af, I-Ak, I-As. ab139365 can inhibit I-A-restricted T cell responses of the H-2b, H-2d, H-2q, H-2u but not H-2f, H-2k, H-2s haplotypes.
  • Tested applications

    Suitable for: Blocking, WB, IP, Functional Studies, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Activated C57BL/6 Mouse spleen cells.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term.
  • Storage buffer

    pH: 7.40
    Constituent: 99% PBS
    Note: PBS is Azide free
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    Purity is > 95% by SDS-PAGE and 0.2 µm filter sterilized.
  • Clonality

    Monoclonal
  • Clone number

    M5/114
  • Isotype

    IgG2b
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab139365 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Blocking Use at an assay dependent concentration. Blocking of T cell proliferative responses
WB Use at an assay dependent concentration. Detects a band of approximately 29 kDa.
IP Use at an assay dependent concentration.
Functional Studies Use at an assay dependent concentration.
Flow Cyt Use a concentration of 2 µg/ml.

ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • 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.
  • Sequence similarities

    Belongs to the MHC class II family.
    Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
  • Cellular localization

    Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network membrane. Endosome membrane. Lysosome membrane. The MHC class II complex transits through a number of intracellular compartments in the endocytic pathway until it reaches the cell membrane for antigen presentation.
  • Information by UniProt
  • Database links

  • Alternative names

    • D6S221E antibody
    • DMA antibody
    • DMB antibody
    • DP beta 1 antibody
    • DP beta 1 chain antibody
    • DP(W4) beta chain antibody
    • DPB 1 antibody
    • DPB1 antibody
    • DPB1_HUMAN antibody
    • DRB antibody
    • H2Ea antibody
    • HLA class II histocompatibility antigen antibody
    • HLA class II histocompatibility antigen DM beta chain antibody
    • HLA class II histocompatibility antigen, DP beta 1 chain antibody
    • HLA class II histocompatibility antigen, DP(W4) beta chain antibody
    • HLA DMB antibody
    • HLA DP1A antibody
    • HLA DPB1 antibody
    • HLA SB alpha chain antibody
    • HLA-A antibody
    • HLA-A histocompatibility type antibody
    • HLA-DP antibody
    • HLA-DP histocompatibility type, beta-1 subunit antibody
    • HLA-DP1B antibody
    • HLA-DPB antibody
    • HLA-DPB1 antibody
    • HLADM antibody
    • HLADP1B antibody
    • HLASB antibody
    • HLASB histocompatibility type antibody
    • Human MHC class II HLA SB alpha antibody
    • LA class II histocompatibility antigen DP alpha 1 chain antibody
    • Major histocompatibility complex class II antibody
    • Major histocompatibility complex class II DP alpha 1 antibody
    • Major histocompatibility complex class II DP beta 1 antibody
    • Major histocompatibility complex, class I, A antibody
    • MHC class II antigen DMB antibody
    • MHC class II antigen DPB1 antibody
    • MHC class II DP3 alpha antibody
    • MHC class II DPA1 antibody
    • MHC class II HLA-DP-beta-1 antibody
    • MHC DPB1 antibody
    • MHC HLA DPB1 antibody
    • PLT1 antibody
    • Primed lymphocyte test 1 antibody
    • RING6 antibody
    • RING7 antibody
    see all

References

This product has been referenced in:

  • Yan A  et al. Partial Depletion of Peripheral M1 Macrophages Reverses Motor Deficits in MPTP-Treated Mouse by Suppressing Neuroinflammation and Dopaminergic Neurodegeneration. Front Aging Neurosci 10:160 (2018). Read more (PubMed: 29922149) »
  • Mori H  et al. Introduction of Zinc-salt Fixation for Effective Detection of Immune Cell-related Markers by Immunohistochemistry. Toxicol Pathol 43:883-9 (2015). Read more (PubMed: 26157038) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH
Permeabilization
No
Specification
Lymph node
Blocking step
Background Sniper (Biocare) as blocking agent for 15 minute(s) · Concentration: 100%
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 16 2016

Answer

Thank you for contacting us.

The laboratory we obtain this clone from claims that antigen retrieval of PFA-fixed samples is not necessary. However, the paper I was sent as validation of this describes a sample preparation that differs a great deal from yours: PFA-fixation of macrophages harvested from cell culture for 2 hours followed by embedding in gelatin, freezing, and preparation of sections from this frozen gelatin, with no subsequent retrieval.

De Souza Leao S, Lang T, Prina E, Hellio R, Antoine JC: Intracellular Leishmania amazonensis amastigotes internalize and degrade MHC class II molecules of their host cells. J Cell Sci. 1995 Oct;108 ( Pt 10):3219-31.

http://jcs.biologists.org/content/108/10/3219.long

A search online using the clone ID, M5/114, and immunohistochemistry and paraformaldehyde as search terms returns many other papers but we do not have access to most journals.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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