Recombinant Anti-HLA-DQA1 antibody [EPR7300] (ab128959)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7300] to HLA-DQA1
- Suitable for: WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-HLA-DQA1 antibody [EPR7300]
See all HLA-DQA1 primary antibodies -
Description
Rabbit monoclonal [EPR7300] to HLA-DQA1 -
Host species
Rabbit -
Specificity
Signals detected in mouse and rat samples are orthologs of HLA.
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Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human HLA-DQA1 aa 200-300 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: Raji cell lysates, mouse and rat spleen lysate. IHC-P: Human liver and tonsil tissues, rat and mouse spleen tissues. IP: Human spleen lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 6.30 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7300 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab128959 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 |
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WB |
1/10000 - 1/50000. Detects a band of approximately 33-35 kDa (predicted molecular weight: 28 kDa).
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IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Heat up to 98 degrees C, below boiling, and then let cool for 10-20 min.
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IP |
Use at an assay dependent concentration.
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Notes |
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WB
1/10000 - 1/50000. Detects a band of approximately 33-35 kDa (predicted molecular weight: 28 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Heat up to 98 degrees C, below boiling, and then let cool for 10-20 min. |
IP
Use at an assay dependent concentration. |
Target
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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 accomodates 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. -
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
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Database links
- Entrez Gene: 100133678 Human
- Entrez Gene: 100507686 Human
- Entrez Gene: 100509457 Human
- Entrez Gene: 3117 Human
- Omim: 146880 Human
- SwissProt: P01909 Human
- Unigene: 387679 Human
- Unigene: 591798 Human
see all -
Alternative names
- CD antibody
- CELIAC1 antibody
- DC 1 alpha chain antibody
see all
Images
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Immunoprecipitation of HLA-DQA1 from 2Human spleen lysate using ab128959 at 1/20 dilution.
Lane 1: Human spleen lysate 10 μg
Lane 2: Human spleen lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab128959 in Human spleen lysate
ab131366 was used as secondary antibody at 1/1000 dilution.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Observed MW at 33-35kDa.
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Immunohistochemical analysis of Paraffin-embedded sections rat spleen tissue labelling HLA-DQA1 with ab128959 at 1/200 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on rat spleen tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
The section was incubated with ab128959 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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Immunohistochemical analysis of Paraffin-embedded sections mouse spleen tissue labelling HLA-DQA1 with ab128959 at 1/200 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on mouse spleen tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
The section was incubated with ab128959 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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Immunohistochemical analysis of Paraffin-embedded sections human tonsil tissue labelling HLA-DQA1 with ab128959 at 1/200 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on human tonsil tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
The section was incubated with ab128959 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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Immunohistochemical analysis of Paraffin-embedded sections human liver tissue labelling HLA-DQA1 with ab128959 at 1/200 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Staining on human liver tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
The section was incubated with ab128959 for 15 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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All lanes : Anti-HLA-DQA1 antibody [EPR7300] (ab128959) at 1/1000 dilution
Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 28 kDa
Observed band size: 33-35 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab128959 has been referenced in 6 publications.
- Asleh K et al. Proteomic analysis of archival breast cancer clinical specimens identifies biological subtypes with distinct survival outcomes. Nat Commun 13:896 (2022). PubMed: 35173148
- Hu Y et al. Differential Expression and Prognostic Correlation of Immune Related Factors Between Right and Left Side Colorectal Cancer. Front Oncol 12:845765 (2022). PubMed: 35936748
- Jeong HO et al. Cellular plasticity and immune microenvironment of malignant pleural effusion are associated with EGFR-TKI resistance in non-small-cell lung carcinoma. iScience 25:105358 (2022). PubMed: 36339256
- Bae JY et al. Evaluation of immune-biomarker expression in high-grade soft-tissue sarcoma: HLA-DQA1 expression as a prognostic marker. Exp Ther Med 20:107 (2020). PubMed: 32989386
- Kanangat S et al. Circulating histocompatibility antigen (HLA) gene products may help differentiate benign from malignant indeterminate pulmonary lesions. Hum Immunol 79:558-563 (2018). WB . PubMed: 29656111
- Kaushansky N et al. Role of a Novel Human Leukocyte Antigen-DQA1*01:02;DRB1*15:01 Mixed Isotype Heterodimer in the Pathogenesis of "Humanized" Multiple Sclerosis-like Disease. J Biol Chem 290:15260-78 (2015). PubMed: 25911099