Recombinant
RabMAb

Recombinant Anti-HLA-DPB1 antibody [EPR11226] - BSA and Azide free (ab215973)

Overview

  • Product name

    Anti-HLA-DPB1 antibody [EPR11226] - BSA and Azide free
    See all HLA-DPB1 primary antibodies
  • Description

    Rabbit monoclonal [EPR11226] to HLA-DPB1 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    corresponding to Human HLA-DPB1.

  • Positive control

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

    Ab215973 is the carrier-free version of ab157210. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab215973 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.
  • Storage buffer

    Constituent: 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 ab215973 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.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

 

WB Use at an assay dependent concentration. Predicted molecular weight: 29 kDa.
  • 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 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 a 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 microenvironment 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

      • beta1 domain MHC class II HLA DPB antibody
      • class II histocompatibility antigen, DP(W4) beta chain antibody
      • class II HLA beta chain antibody
      • DP beta 1 chain antibody
      • DP(W4) beta chain antibody
      • DPB1 antibody
      • DPB1_HUMAN antibody
      • HLA class II histocompatibility antigen antibody
      • HLA class II histocompatibility antigen, DP beta 1 chain antibody
      • HLA class II histocompatibility antigen, DP(W4) beta chain antibody
      • HLA DP14-beta chain antibody
      • HLA-DP antibody
      • HLA-DP histocompatibility type, beta-1 subunit antibody
      • HLA-DP1B antibody
      • HLA-DPB antibody
      • HLA-DPB1 antibody
      • major histocompatibility complex class II antigen beta chain antibody
      • major histocompatibility complex, class II, DP beta 1 antibody
      • MHC class II antigen beta chain antibody
      • MHC class II antigen DP beta 1 chain antibody
      • MHC class II antigen DPB1 antibody
      • MHC class II antigen DPbeta1 antibody
      • MHC class II HLA-DP-beta-1 antibody
      • MHC class II HLA-DRB1 antibody
      • MHC HLA DPB1 antibody
      see all

    Images

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

    • 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, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

    • Unpurified ab157210 showing negative staining in Human heart.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Unpurified ab157210 showing negative staining in Human normal brain.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Unpurified ab157210 showing positiveve staining in human normal colon.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunofluorescent analysis of Jurkat cells labeling MHC Class II with unpurified ab157210 at 1/50 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab157210).

    References

    This product has been referenced in:

    • Arebro J  et al. Antigen-presenting epithelial cells can play a pivotal role in airway allergy. J Allergy Clin Immunol N/A:N/A (2015). IF, IHC ; Human . Read more (PubMed: 26560042) »
    See 1 Publication for this product

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