Product nameHuman ATF6 knockout HeLa cell lysate
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 17 bp deletion in exon6 and 25 bp deletion in exon6 and 4 bp deletion in exon6.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo 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.
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: After reconstitution, store the lysate at -80°C.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab261973 - Human ATF6 knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255929 - Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
FunctionTranscription factor that acts during endoplasmic reticulum stress by activating unfolded protein response target genes. Binds DNA on the 5'-CCAC[GA]-3'half of the ER stress response element (ERSE) (5'-CCAAT-N(9)-CCAC[GA]-3') and of ERSE II (5'-ATTGG-N-CCACG-3'). Binding to ERSE requires binding of NF-Y to ERSE. Could also be involved in activation of transcription by the serum response factor.
Sequence similaritiesBelongs to the bZIP family. ATF subfamily.
Contains 1 bZIP domain.
DomainThe basic domain functions as a nuclear localization signal.
The basic leucine-zipper domain is sufficient for association with the NF-Y trimer and binding to ERSE.
modificationsDuring unfolded protein response an approximative 50 kDa fragment containing the cytoplasmic transcription factor domain is released by proteolysis. The cleavage seems to be performed sequentially by site-1 and site-2 proteases.
N-glycosylated. The glycosylation status may serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the unfolded protein response (UPR).
Phosphorylated in vitro by MAPK14/P38MAPK.
Cellular localizationEndoplasmic reticulum membrane and Nucleus. Under ER stress the cleaved N-terminal cytoplasmic domain translocates into the nucleus.
- Information by UniProt
- Activating transcription factor 6
- Activating transcription factor 6 alpha
- ATF 6
KO cell lines
Our Abpromise guarantee covers the use of ab256841 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 74 kDa.|
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ATF6 knockout HeLa cell lysate (20 µg)
Lane 3: Wild-type HAP1 cell lysate (20 µg)
Lane 4: ATF6 knockout HAP1 cell lysate (20 µg)
ab83504 Anti-ATF6 antibody was shown to specifically react with ATF6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261800 (knockout cell lysate ab256841) was used. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. ab83504 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Allele-1: 25 bp deletion in exon6
Allele-2: 17 bp deletion in exon6
Allele-3: 4 bp deletion in exon6
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.
ab256841 has not yet been referenced specifically in any publications.