Recombinant
RabMAb

Anti-HLA-DR antibody [EPR3692] (ab92511)

Overview

  • Product name
    Anti-HLA-DR antibody [EPR3692]
    See all HLA-DR primary antibodies
  • Description
    Rabbit monoclonal [EPR3692] to HLA-DR
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Rat, Human
    Does not react with: Mouse
  • Immunogen

    corresponding to Human HLA-DR aa 150-250.

  • Positive control
    • WB: Raji, Human spleen and Hu T-78 cell lysates IHC-P: Human tonsil and Human kidney tissues
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

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.
  • Dissociation constant (KD)
    KD = 1.67 x 10 -10 M Learn more about KD
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR3692
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab92511 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
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/5000 - 1/10000. Predicted molecular weight: 29 kDa.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
  • Application notes
    Is unsuitable for IP.
  • 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 accommodates 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.
    • Post-translational
      modifications
      Ubiquitinated by MARCH1 or MARCH8 at Lys-244 leading to down-regulation of MHC class II. When associated with ubiquitination of the beta subunit of HLA-DR: HLA-DRB4 'Lys-254', the down-regulation of MHC class II may be highly effective.
    • Cellular localization
      Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus > trans-Golgi network membrane. Endosome membrane. Lysosome membrane. Late endosome 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
      • DR alpha chain antibody
      • DR alpha chain precursor antibody
      • DRA_HUMAN antibody
      • DRB1 antibody
      • DRB4 antibody
      • Histocompatibility antigen HLA DR alpha antibody
      • HLA class II histocompatibility antigen antibody
      • HLA class II histocompatibility antigen DR alpha chain antibody
      • HLA DR1B antibody
      • HLA DR3B antibody
      • HLA DRA antibody
      • HLA DRA1 antibody
      • HLA DRB1 antibody
      • HLA DRB3 antibody
      • HLA DRB4 antibody
      • HLA DRB5 antibody
      • HLA-DRA antibody
      • HLADR4B antibody
      • HLADRA1 antibody
      • HLADRB antibody
      • Major histocompatibility complex class II DR alpha antibody
      • Major histocompatibility complex class II DR beta 1 antibody
      • Major histocompatibility complex class II DR beta 3 antibody
      • Major histocompatibility complex class II DR beta 4 antibody
      • Major histocompatibility complex class II DR beta 5 antibody
      • MGC117330 antibody
      • MHC cell surface glycoprotein antibody
      • MHC class II antigen DRA antibody
      • MHC II antibody
      • MLRW antibody
      see all

    Anti-HLA-DR antibody [EPR3692] images

    • ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

      Negative control 1: PBS in place of primary antibody.

    • ab92511 staining HLA-DR in HuT-78 (human Sezary syndrome) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab92511 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

      Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
      Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

    • All lanes : Anti-HLA-DR antibody [EPR3692] (ab92511) at 1/10000 dilution

      Lane 1 : Raji (human Burkitt's lymphoma) whole cell lysate
      Lane 2 : Ramos (human Burkitt's lymphoma) whole cell lysate
      Lane 3 : Human tonsil

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

      Predicted band size : 29 kDa

      Blocking buffer: 5% NFDM/TBST

      Diluting buffer: 5% NFDM/TBST

    • Overlay histogram showing Raji cells stained with ab92511 (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 (ab92511, 1/50) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.

    • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling HLA DR with ab92511 at a dilution of 1/1000. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as the secondary antibody, at a dilution of 1/500. Counter stained with hematoxylin.

    • All lanes : Anti-HLA-DR antibody [EPR3692] (ab92511) at 1/5000 dilution

      Lane 1 : Raji cell lysate
      Lane 2 : Human spleen lysate
      Lane 3 : Hu T-78 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 29 kDa
    • ab92511 showing positive staining in Normal liver vessels tissue.

    • ab92511 showing positive staining in Normal skin vessels tissue.

    • ab92511 showing positive staining in Endometrial carcinoma vessels tissue.

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References for Anti-HLA-DR antibody [EPR3692] (ab92511)

    This product has been referenced in:
    • Watanabe N  et al. A cell-based high-throughput screening assay system for inhibitor compounds of antigen presentation by HLA class II molecule. Sci Rep 7:6798 (2017). Read more (PubMed: 28754892) »
    • Mastropasqua R  et al. Corneoscleral limbus in glaucoma patients: in vivo confocal microscopy and immunocytological study. Invest Ophthalmol Vis Sci 56:2050-8 (2015). Read more (PubMed: 25744981) »

    See all 6 Publications for this product

    Product Wall

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Liver)
    Specification
    Liver
    Fixative
    Formaldehyde
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C
    Username

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    Verified customer

    Submitted Jul 08 2011

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

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