HLA-DP Recombinant Rabbit Monoclonal Antibody [PD00-70]
Catalog# HA721247
HLA-DP Recombinant Rabbit Monoclonal Antibody [PD00-70]
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WB
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IF-Cell
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FC
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IHC-P
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Human
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unconjugated
Overview
Product Name
HLA-DP Recombinant Rabbit Monoclonal Antibody [PD00-70]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within C terminal Human HLA-DP.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, FC, IHC-P
Molecular Weight
Predicted band size: 29 kDa
Positive Control
Raji cell lysate, Daudi cell lysate, Ramos cell lysate, Daudi.
Conjugation
unconjugated
Clone Number
PD00-70
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IHC-P
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1: 200
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IF-Cell
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1:100
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FC
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1:500-1:1,000
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.
Background References
1. Xia Y et al. The association of HLA-DP loci with autoimmune diabetes in Chinese. Diabetes Res Clin Pract. 2021 Mar
2. Daniëls L et al. The role of HLA-DP mismatches and donor specific HLA-DP antibodies in kidney transplantation: a case series. Transpl Immunol. 2021 Apr
Subcellular Location
Cell membrane, Endoplasmic reticulum membrane, Golgi apparatus, Endosome membrane, Lysosome membrane.
UNIPROT
Synonyms
DP(W3) antibody
DP(W4) antibody
HLA class II histocompatibility antigen DP alpha 1 chain antibody
HLA class II histocompatibility antigen DP alpha chain antibody
HLA DP1A antibody
HLA DPA 1 antibody
HLA SB alpha chain antibody
HLADP antibody
HLADPA1 antibody
HLASB antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of HLA-DP on different lysates with Rabbit anti-HLA-DP antibody (HA721247) at 1/5,000 dilution.
Lane 1: Raji cell lysate (20 µg/Lane)
Lane 2: Daudi cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (negative) (20 µg/Lane)
Lane 4: Ramos cell lysate (20 µg/Lane)
Predicted band size: 29 kDa
Observed band size: 35 kDa
Exposure time: 4 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721247) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-HLA-DP antibody (HA721247) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721247) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue with Rabbit anti-HLA-DP antibody (HA721247) at 1/500 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721247) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunocytochemistry analysis of Daudi cells labeling HLA-DP with Rabbit anti-HLA-DP antibody (HA721247) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HLA-DP antibody (HA721247) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of Daudi cells labeling HLA-DP.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721247, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"