Product nameHuman AES (Amino-terminal enhancer of split) knockout HCT116 cell lysate
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHCT116
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon2.
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 ab263593 - Human AES knockout HCT116 cell lysate (Lyophilized) 1 x 100µg ab255555 - Human wild-type HCT116 cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 10, 11 D13S317: 10, 12 D7S820: 11, 12 D16S539: 11, 13 vWA: 17, 22 TH01: 8,9 TPOX: 8, 9 CSF1PO: 7, 10
FunctionTranscriptional corepressor. Acts as dominant repressor towards other family members. Inhibits NF-kappa-B-regulated gene expression. May be required for the initiation and maintenance of the differentiated state. Essential for the transcriptional repressor activity of SIX3 during retina and lens development.
Tissue specificityFound predominantly in muscle, heart and Placenta. In fetal tissues, abundantly expressed in the heart, lung, kidney, brain and liver.
Sequence similaritiesBelongs to the WD repeat Groucho/TLE family.
DomainLacks the C-terminal WD repeats.
modificationsUbiquitinated by XIAP/BIRC4.
- Information by UniProt
- Aes 1
- Aes 2
KO cell lines
Our Abpromise guarantee covers the use of ab257818 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: 21 kDa.|
Lane 1:Wild-type HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 2:AES knockout HCT 116 (Human colorectal carcinoma cell line) whole cell lysate (20 ug)
Lane 3:HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 4:PC3 (Human prostate adenocarcinoma cell line) whole cell lysate (20 ug)
ab137060 was shown to specifically react with Amino-terminal enhancer of split/AES in wild-type HCT cells. Loss of signal was observed when knockout cell line ab266886 (knockout cell lysate ab257818) was used. Wild-type and Amino-terminal enhancer of split/AES knockout samples were subjected to SDS-PAGE. ab137060 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 1000 dilution 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.
Homozygous: 1 bp deletion in exon2
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.
ab257818 has not yet been referenced specifically in any publications.