Human HLA-E (HLA E) knockout A549 cell lysate (ab258452)
Overview
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Product name
Human HLA-E (HLA E) knockout A549 cell lysate
See all HLA E kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2. -
Passage number
Knockout validation
Sanger Sequencing, 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.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262418 - Human HLA-E knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
Target
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Relevance
HLA E 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. HLA E binds a restricted subset of peptides derived from the leader peptides of other class I molecules. -
Cellular localization
Membrane; Single-pass type I membrane protein -
Alternative names
- HLA class I histocompatibility antigen E alpha chain
- EA1.2
- EA2.1
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262418 - Human HLA-E knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
Images
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Western blot - Human HLA-E (HLA E) knockout A549 cell lysate (ab258452)
Lane 1:Wild-type A549 cell lysate (20 ug)
Lane 2:HLA-E knockout A549 cell lysate (20 ug)
Lane 3:THP-1 cell lysate (20 ug)
Lane 4:Jurkat cell lysate (20 ug)ab2216 was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267080 (knockout cell lysate ab258452) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Sanger Sequencing - Human HLA-E knockout A549 cell lysate (ab258452)Homozygous: 1 bp insertion in exon2
