Recombinant
RabMAb

Recombinant Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free (ab215985)

Overview

  • Product name

    Anti-HLA-DR antibody [EPR3692] - Low endotoxin, Azide free
    See all HLA-DR primary antibodies
  • Description

    Rabbit monoclonal [EPR3692] to HLA-DR - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, IHC-P, WBmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse
  • Immunogen

    Synthetic peptide within Human HLA-DR aa 150-250. The exact sequence is proprietary.

  • Positive control

    • WB: Raji, Human spleen and Hu T-78 cell lysates IHC-P: Human tonsil, spleen, endometrial cancer, skin and kidney tissues. Flow cyt: Raji cells. ICC/IF: Hut-78 cells.
  • General notes

    ab215985 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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.

    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 long term. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 1.67 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3692
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab215985 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
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Predicted molecular weight: 29 kDa.
  • 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

      • DASS-397D15.1 antibody
      • DR alpha chain antibody
      • DR alpha chain precursor antibody
      • DRA_HUMAN antibody
      • DRB1 antibody
      • DRB4 antibody
      • FLJ51114 antibody
      • Histocompatibility antigen HLA DR alpha 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-DR histocompatibility type 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
      • OTTHUMP00000029406 antibody
      • OTTHUMP00000029407 antibody
      see all

    Images

    • Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-HLA-DR antibody [EPR3692]. ab215985 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.

      Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • 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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • ab92511 showing positive staining in Normal liver vessels tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • ab92511 showing positive staining in Normal skin vessels tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • ab92511 showing positive staining in Endometrial carcinoma vessels tissue.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92511).

    • This IHC data was generated using the same anti-HLA DR antibody clone, EPR3692, in a different buffer formulation (cat# ab92511).

      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.

    References

    This product has been referenced in:

    • 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) »
    • Steain M  et al. Analysis of T cell responses during active varicella-zoster virus reactivation in human ganglia. J Virol 88:2704-16 (2014). Read more (PubMed: 24352459) »
    See all 3 Publications for this product

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