Recombinant Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22172] to HLA Class 1 ABC
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Reacts with: Human
Overview
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Product name
Anti-HLA Class 1 ABC antibody [EPR22172]
See all HLA Class 1 ABC primary antibodies -
Description
Rabbit monoclonal [EPR22172] to HLA Class 1 ABC -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment within Human HLA Class 1 ABC aa 1-300. The exact sequence is proprietary.
Database link: P01889 -
Positive control
- ICC/IF: THP-1 and Raji cells. Flow Cyt: Raji and THP-1 cells. IHC-P: Human colonic carcinoma, stomach and liver tissue. WB: Raji, THP-1, HL-60, Jurkat, HepG2, TF-1 and Ramos whole cell lysate. Human tonsil. Recombinant human HLA-A protein (aa24-aa365). Recombinant human HLA-C protein (aa25-aa308). IP: HLAB in THP-1 whole cell 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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
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.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22172 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry reagents
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab225636 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 | 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). | |
IHC-P | 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
ICC/IF | 1/250. | |
Flow Cyt | 1/600. | |
IP | 1/30. |
Target
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Relevance
HLA-ABC antigen is present on most nucleated cells of humans and Old World monkeys. HLA CLass I is involved in the presentation of foreign antigens to the immune system. -
Cellular localization
Cell Membrane; Single-pass type I membrane protein -
Database links
- Entrez Gene: 3105 Human
- Entrez Gene: 3106 Human
- Entrez Gene: 3107 Human
- SwissProt: P01889 Human
- SwissProt: P30443 Human
- SwissProt: P30499 Human
- Unigene: 181244 Human
- Unigene: 654404 Human
see all -
Alternative names
- Antigen Presenting Molecule antibody
- Cw*1701 antibody
- D6S204 antibody
see all
Images
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All lanes : Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636) at 1/1000 dilution
Lane 1 : Recombinant human HLA-A protein (aa24-aa365) 10ng
Lane 3 : Recombinant human HLA-C protein (aa25-aa308) 10ng
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 40 kDaThis antibody has cross reaction with human HLA-A and HLA-C.# Recombinant human HLA-A protein was purchased from abnova company (H00003105-P01).
Recombinant human HLA-C protein was purchased from novusbio company (NBP2-23210).
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Immunofluorescent analysis of 100% methanol-fixed THP-1 (human monocytic leukemia cell line) cells labeling HLAB with ab225636 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in THP-1 cells. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
The negative control is the secondary antibody only.Methanol is recommended for fixation as weak signal was observed using 4% PFA.
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Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cells labeling HLAB with ab225636 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on lymph nodule of human stomach (PMID:25294906) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
All lanes : Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636) at 1/1000 dilution
Lane 1 : Raji (human Burkitt's lymphoma cell line), whole cell lysate
Lane 2 : THP-1 (human monocytic leukemia cell line), whole cell lysate
Lane 3 : HL-60 (human promyelocytic leukemia cell line), whole cell lysate
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood), whole cell lysate
Lane 5 : HepG2 (human liver hepatocellular carcinoma cell line), whole cell lysate
Lane 6 : TF-1 (human bone marrow erythroleukemia cell line), whole cell lysate
Lane 7 : Ramos (human Burkitt's lymphoma cell line), whole cell lysate
Lane 8 : Human tonsil
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
Exposure time: 26 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Weak reactive bands of undetermined proteins are observed in addition to the target band.
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HLAB was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia cell line) whole cell lysate with ab225636 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab225636 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: THP-1 whole cell lysate 10 µg (Input).
Lane 2: ab225636 IP in THP-1 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab225636 in THP-1 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 seconds. -
Immunofluorescent analysis of 100% methanol-fixed Raji (human Burkitt's lymphoma cell line) cells labeling HLAB with ab225636 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Raji cells. The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).
The negative control is the secondary antibody only.Methanol is recommended for fixation as weak signal was observed using 4% PFA.
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Flow cytometric analysis of THP-1 (human monocytic leukemia cell line) cell line labeling HLAB with ab225636 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on hepatic sinus endothelium of human liver (PMID: 26963506) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HLA Class 1 ABC antibody [EPR22172] (ab225636)
Immunohistochemical analysis of paraffin-embedded human colonic carcinoma tissue labeling HLAB with ab225636 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on tumor cells of human colonic carcinoma (PMID: 26310568, PMID: 17014712) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Protocols
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.
Datasheets and documents
Certificate of Compliance
References (0)
ab225636 has not yet been referenced specifically in any publications.