Product nameHuman MAPK12 knockout HEK293T cell lysate
See all MAPK 12 kits
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHEK293T
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 4 and 5 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 ab261123 - Human MAPK12 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
FunctionResponds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating downstream targets. Plays a role in myoblast differentiation and also in the down-regulation of cyclin D1 in response to hypoxia in adrenal cells suggesting MAPK12 may inhibit cell proliferation while promoting differentiation.
Tissue specificityHighly expressed in skeletal muscle and heart.
Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain.
DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
modificationsDually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme.
Cellular localizationCytoplasm. Mitochondrion. Mitochondrial when associated with SH3BP5.
- Information by UniProt
- ERK 6
KO cell lines
Our Abpromise guarantee covers the use of ab258041 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: 42 kDa.|
Lane 1:Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2:MAPK12 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3:Human skeletal muscle tissue lysate (20 ug)
ab205926 was shown to specifically react with MAPK 12 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266280 (knockout cell lysate ab258041) was used. Wild-type and MAPK 12 knockout samples were subjected to SDS-PAGE. ab205926 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.
Allele-1: 5 bp deletion in exon 4
Allele-2: 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.
ab258041 has not yet been referenced specifically in any publications.