Recombinant
RabMAb

Anti-MHC Class II antibody [EPR11226] (ab157210)

Overview

  • Product name
    Anti-MHC Class II antibody [EPR11226]
    See all MHC Class II primary antibodies
  • Description
    Rabbit monoclonal [EPR11226] to MHC Class II
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MHC Class II.

  • Positive control
    • Human fetal thymus and Human tonsil lysates; Human tonsil tissue; Jurkat cells; Immunoprecipitation pellet from fetal thymus lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR11226
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab157210 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
WB 1/10000 - 1/50000. Predicted molecular weight: 29 kDa.
IHC-P 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified, use 1/100 - 1/250.

ICC/IF 1/50 - 1/250.
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt.
  • 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

    Images

    • Immunohistochemical staining of paraffin embedded human tonsil with purified ab157210 at a working dilution of 1/2500. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunofluorescent analysis of Jurkat cells labeling MHC Class II with unpurified ab157210 at 1/50 dilution.

    • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling MHC Class II with unpurified ab157210 at 1/100 dilution.

    • All lanes : Anti-MHC Class II antibody [EPR11226] (ab157210) at 1/20000 dilution (purified)

      Lane 1 : Human fetal thymus lysate
      Lane 2 : Human tonsil lysate
      Lane 3 : Human fetal spleen lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 29 kDa
      Observed band size: 29 kDa



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • ab157210 (purified) at 1/70 immunoprecipitating MHC Class II in 10 μg Daudi cell lysate (Lanes 1 and 2, observed at 29 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
    • Immunofluorescence staining of Raji cells with purified ab157210 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab157210 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Immunohistochemical staining of paraffin embedded human skeletal muscle with purified ab157210 at a working dilution of 1/2500. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Unpurified ab157210 showing positiveve staining in human normal colon.

    • Unpurified ab157210 showing negative staining in Human heart.

    • Unpurified ab157210 showing negative staining in Human normal brain.

    • All lanes : Anti-MHC Class II antibody [EPR11226] (ab157210) at 1/10000 dilution (Unpurified)

      Lane 1 : Human fetal thymus cell lysate
      Lane 2 : Human tonsil cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 29 kDa

    • Anti-MHC Class II antibody [EPR11226] (ab157210) at 1/10000 dilution (Unpurified) + Immunoprecipitation pellet from Human fetal thymus lysate at 10 µg

      Predicted band size: 29 kDa

    References

    This product has been referenced in:
    • Hong D  et al. Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection. Stem Cell Res Ther 9:49 (2018). WB . Read more (PubMed: 29482598) »
    • Han S  et al. LPS alters the immuno-phenotype of glioma and glioma stem-like cells and induces in vivo antitumor immunity via TLR4. J Exp Clin Cancer Res 36:83 (2017). Read more (PubMed: 28641579) »

    See all 3 Publications for this product

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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