Product nameHuman CAPN2 (Calpain 2) knockout HEK293T cell lysate
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHEK293T
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 17 bp deletion in exon 4 and 1 bp insertion in exon 4 and 47 bp deletion in exon 4 and 8 bp deletion in exon 4.
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, 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 applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab260215 - Human CAPN2 knockout HEK293T cell lysate (Lyophilized) 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
FunctionCalcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
Sequence similaritiesBelongs to the peptidase C2 family.
Contains 1 calpain catalytic domain.
Contains 3 EF-hand domains.
Cellular localizationCytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
- Information by UniProt
- Calcium activated neutral proteinase
- Calcium activated neutral proteinase 2
- Calcium-activated neutral protease 2, catalytic subunit
KO cell lines
Our Abpromise guarantee covers the use of ab257379 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: 80 kDa.|
Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:CAPN2 knockout HEK293T cell lysate (20 ug)
Lane 3:A431 cell lysate (20 ug)
Lane 4:LNCaP cell lysate (20 ug)
ab126600 was shown to specifically react with Calpain 2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266628 (knockout cell lysate ab257379) was used. Wild-type and Calpain 2 knockout samples were subjected to SDS-PAGE. ab126600 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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.
Allele-1: 47 bp deletion in exon 4
Allele-2: 17 bp deletion in exon 4
Allele-3: 8 bp deletion in exon 4
Allele-4: 1 bp insertion in exon 4
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.
ab257379 has not yet been referenced specifically in any publications.