Overview

  • Product nameAnti-MHC Class II antibodySee all MHC Class II primary antibodies ...
  • Description
    Mouse monoclonal to MHC Class II
  • Tested applicationsIHC-P, ICC/IF, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Human, Recombinant Fragment
  • Immunogen

    Recombinant full length protein, corresponding to amino acids 1-259 of Human MHC Class II beta chain HLA-DPB1

  • Positive control
    • This antibody gave a positive result when used in the following formaldehyde fixed cell lines: A431

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: None
    PBS, pH 7.2
  • Concentration information loading...
  • PurityProtein G purified
  • Clonality Monoclonal
  • IsotypeIgG2a
  • Light chain typekappa
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab55152 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
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 - 5 µg/ml.
This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein.
Flow Cyt Use 0.5µg for 106 cells. (Also see PMID: 18941249)

Target

  • FunctionBinds 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 similaritiesBelongs to the MHC class II family.
    Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
  • Cellular localizationCell 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 DMB antibody
    • HLA DPB1 antibody
    • HLA-DP antibody
    • HLA-DP1B antibody
    • HLA-DPB antibody
    • HLA-DPB1 antibody
    • HLADM antibody
    • HLADP1B antibody
    • Major histocompatibility complex class II antibody
    • Major histocompatibility complex class II DP beta 1 antibody
    • MHC class II antigen DMB antibody
    • MHC class II antigen DPB1 antibody
    • MHC DPB1 antibody
    • RING6 antibody
    • RING7 antibody
    see all

Anti-MHC Class II antibody images

  • ICC/IF image of ab55152 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab55152 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • MHC Class II antibody (ab55152) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human lymph node.
  • Western blot against tagged recombinant protein immunogen using ab55152 MHC Class II antibody at 1ug/ml. Predicted band size of immunogen is 54 kDa
  • Overlay histogram showing Raji cells stained with ab55152 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab55152, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-MHC Class II antibody (ab55152)

This product has been referenced in:
  • Schmidt K  et al. Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor 3 Inhibits Gamma Interferon and Major Histocompatibility Complex Class II Expression. J Virol 85:4530-7 (2011). WB ; Human . Read more (PubMed: 21345951) »
  • Liu YH  et al. Up-regulation of vascular endothelial growth factor-D expression in clear cell renal cell carcinoma by CD74: a critical role in cancer cell tumorigenesis. J Immunol 181:6584-94 (2008). Flow Cyt ; Human . Read more (PubMed: 18941249) »

See all 2 Publications for this product

Product Wall

The red is Wheat germ agglutinin (WGA). A widely used lectin in cell biology which has been conjugated to Alexa Fluor 594. WGA binds to sialic acid and N-acetylglucosaminyl residues.

Thank you, the images all arrived. I think you are correct, the TIFF files were probably too big.

Have you been able to obtain good stains for other proteins using this protocol?

I think the protocol is correct as it is. A custome...

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Thank you for contacting us.


I have searched our catalog and unfortunately we do not currently have any anti-MHC Class II antibodies which have been tested in hamster. Looking further at the hamster protein against the mouse, rat and ...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab116378. To check the status of the order please contact our Custome...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (bowel)
Specification bowel
Fixative Formaldehyde
Antigen retrieval step Enzymatic
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Aug 03 2011

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