Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP229] to HLA-DPB1
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
- Reacts with: Human
Product nameAnti-HLA-DPB1 antibody [SP229]
See all HLA-DPB1 primary antibodies
DescriptionRabbit monoclonal [SP229] to HLA-DPB1
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
Species reactivityReacts with: Human
Synthetic peptide within Human HLA-DPB1 aa 200 to the C-terminus. The exact sequence is proprietary.
Database link: P04440
- WB: Ramos, Raji and HeLa cell lysates; human spleen tissue lysate IHC-P: Human tonsil tissue; Flow Cyt: Ramos cells; ICC: Ramos cells.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped 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 bufferpH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Concentration information loading...
PurityProtein A/G purified
Purification notesPurified from TCS by Protein A/G.
Our Abpromise guarantee covers the use of ab227676 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 0.11 µg/ml. Predicted molecular weight: 29 kDa.|
|IHC-P||1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Primary antibody incubation for 10 minutes at room temperature.
|Flow Cyt||1/20 - 1/400.
Primary antibody incubation for 10 minutes at 4°C.
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 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 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
- beta1 domain MHC class II HLA DPB antibody
- class II histocompatibility antigen, DP(W4) beta chain antibody
- class II HLA beta chain antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling HLA-DPB1 with ab227676 at 1/100 dilution (1.10 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
All lanes : Anti-HLA-DPB1 antibody [SP229] (ab227676) at 0.11 µg/ml
Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte)
Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte)
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell)
Lane 4 : Human spleen tissue lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/ml
Predicted band size: 29 kDa
Observed band size: 29 kDa
Blocking/diluting Buffer and concentration: 5% NFDM /TBST
Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) cells labeling HLA-DPB1 with purified ab227676 at 1/10 (10 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling HLA-DPB1 with purified ab227676 at 1/20 dilution (5.5 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.
Formalin-fixed, paraffin-embedded human tonsil tissue stained for HLA-DPB1 using ab227676 at 1/100 dilution in immunohistochemical analysis.
Flow cytometric analysis of Ramos (human Burkitt's lymphoma cell line) cell line labeling HLA-DPB1 with ab227676 at 1/400 dilution (green) compared with a negative control of rabbit IgG (blue).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab227676 has not yet been referenced specifically in any publications.