Human HLA-A knockout A-431 cell lysate (ab261703)
Overview
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Product name
Human HLA-A knockout A-431 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 17 bp deletion; Frameshift = 99.8% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280453 - Human HLA-A knockout A431 cell lysate 1 x 100µg ab263973 - Human wild-type A-431 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
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
- HLA class I histocompatibility antigen, A 1 alpha chain
- HLAA
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261703 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 |
Use at an assay dependent concentration. Predicted molecular weight: 41 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 41 kDa. |
Images
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Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2: EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab86597 observed at 40 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab86597 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab86597 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. -
Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 2: HLA A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 3: A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab52922 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab52922 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab52922 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 developed 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. -
X = 17 bp deletion
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
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SDS download
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Datasheet download
References (0)
ab261703 has not yet been referenced specifically in any publications.