Product nameHuman CAMK2D (CaMKII delta) knockout HEK293T cell lysate
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHEK293T
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 14 bp deletion in exon5 and 1 bp insertion in exon5.
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 ab262074 - Human CAMK2D 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
FunctionCaM-kinase II (CAMK2) is a prominent kinase in the central nervous system that may function in long-term potentiation and neurotransmitter release.
Tissue specificityExpressed in cardiac muscle and skeletal muscle. Isoform Delta 3, isoform Delta 2, isoform Delta 8 and isoform Delta 9 are expressed in cardiac muscle. Isoform Delta 11 is expressed in skeletal muscle.
Sequence similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.
Contains 1 protein kinase domain.
- Information by UniProt
- Calcium / calmodulin dependent protein kinase 2 delta
- Calcium / calmodulin dependent protein kinase II delta
- calcium/calmodulin-dependent protein kinase (CaM kinase) II delta
KO cell lines
Our Abpromise guarantee covers the use of ab257376 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: 56 kDa.|
Lane 1: Wild-type HEK-293T cell lysate (20µg)
Lane 2: CAMK2D knockout HEK-293T cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab181052 observed at 50 kDa. Red - loading control ab8245 observed at 37 kDa.
ab181052 Anti-CaMKII delta antibody [EPR13095] was shown to specifically react with CaM-kinase II in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab267322 (knockout cell lysate ab257376) was used. Wild-type and CaM-kinase II knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab181052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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: 14 bp deletion in exon5
Allele-2: 1 bp insertion in exon5
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.
ab257376 has not yet been referenced specifically in any publications.