Recombinant HRP Anti-HLA A antibody [EP1395Y] (ab199555)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HLA A antibody [EP1395Y] (ab199555)](https://www.abcam.com/ps/products/199/ab199555/Images/ab199555-249415-IHCAP22906761.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HLA A antibody [EP1395Y] (ab199555)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- HRP Rabbit monoclonal [EP1395Y] to HLA A
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
Overview
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Product name
HRP Anti-HLA A antibody [EP1395Y]
See all HLA A primary antibodies -
Description
HRP Rabbit monoclonal [EP1395Y] to HLA A -
Host species
Rabbit -
Conjugation
HRP -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human HLA A aa 50-150. The exact sequence is proprietary.
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Positive control
- WB: A431, A549, Jurkat and Raji whole cell lysate. IHC-P: Normal human tonsil tissue sections.
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General notes
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. Avoid freeze / thaw cycle. Store In the Dark. -
Storage buffer
pH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: PBS, 30% Glycerol, 1% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1395Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-HLA A antibody [EP1395Y] (ab199780)
- Alexa Fluor® 647 Anti-HLA A antibody [EP1395Y] (ab199837)
- Alexa Fluor® 594 Anti-HLA A antibody [EP1395Y] (ab207872)
- Anti-HLA A antibody [EP1395Y] - BSA and Azide free (ab216653)
- Anti-HLA A antibody [EP1395Y] - Low endotoxin, Azide free (ab246691)
- Anti-HLA A antibody [EP1395Y] (ab52922)
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Isotype control
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Positive Controls
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Related Products
Applications
Our Abpromise guarantee covers the use of ab199555 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 |
|---|---|---|
| WB | 1/10000 - 1/50000. Predicted molecular weight: 41 kDa. | |
| IHC-P | 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Relevance
HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen. They are expressed in nearly all cells. The heavy chain is approximately 45 kDa and its gene contains 8 exons. Exon 1 encodes the leader peptide, exons 2 and 3 encode the alpha1 and alpha2 domains, which both bind the peptide, exon 4 encodes the alpha3 domain, exon 5 encodes the transmembrane region, and exons 6 and 7 encode the cytoplasmic tail. Polymorphisms within exon 2 and exon 3 are responsible for the peptide binding specificity of each class one molecule. Typing for these polymorphisms is routinely done for bone marrow and kidney transplantation. Hundreds of HLA-A alleles have been described. -
Alternative names
- Antigen presenting molecule antibody
- HLA class I histocompatibility antigen, A 1 alpha chain antibody
- HLAA antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-HLA A antibody [EP1395Y] (ab199555)
IHC image of HLA A staining in a section of formalin-fixed paraffin-embedded normal human tonsil*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab199555, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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![Western blot - HRP Anti-HLA A antibody [EP1395Y] (ab199555)](https://www.abcam.com/ps/products/199/ab199555/Images/ab199555-395840-anti-hla-a-antibody-ep1395y-hrp-western-blot-a431-wt-a431-hlaa-knockout-a549-jurkat2.jpg)
Western blot - HRP Anti-HLA A antibody [EP1395Y] (ab199555)All lanes : HRP Anti-HLA A antibody [EP1395Y] (ab199555) at 1/10000 dilution
Lane 1 : Wild-type A431 cell lysate
Lane 2 : HLA-A knockout A431 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 41 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab199555 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab199555 was shown to react with HLA A (HRP) in wild-type A431 cells in western blot. Loss of signal was observed when HLA-A knockout sample was used. Wild-type and HLA-A knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab199555 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - HRP Anti-HLA A antibody [EP1395Y] (ab199555)](https://www.abcam.com/ps/products/199/ab199555/Images/ab199555-246522-WBab1995551.jpg)
Western blot - HRP Anti-HLA A antibody [EP1395Y] (ab199555)HRP Anti-HLA A antibody [EP1395Y] (ab199555) at 1/5000 dilution + Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 41 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab199555 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
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![HRP Anti-HLA A antibody [EP1395Y] (ab199555)](https://www.abcam.com/ps/products/199/ab199555/Images/ab199555-5-benefits-of-recombinant-antibodies.png)
HRP Anti-HLA A antibody [EP1395Y] (ab199555)
Protocols
References (0)
ab199555 has not yet been referenced specifically in any publications.
