Product nameHuman GK (Glycerol kinase) knockout HEK293T cell lysate
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineHEK293T
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
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.
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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 ab262988 - Human GK knockout HEK293T cell lysate (Lyophilized) 1 x 100µg ab255553 - Human Wild Type HEK293T cell lysate (Lyophilized) 1 x 100µg
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
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
FunctionKey enzyme in the regulation of glycerol uptake and metabolism.
Tissue specificityHighly expressed in the liver, kidney and testis. Isoform 2 and isoform 3 are expressed specifically in testis and fetal liver, but not in the adult liver.
PathwayPolyol metabolism; glycerol degradation via glycerol kinase pathway; sn-glycerol 3-phosphate from glycerol: step 1/1.
Involvement in diseaseDefects in GK are the cause of GK deficiency (GKD) [MIM:307030]. This disease can be either symptomatic with episodic metabolic and CNS decompensation or asymptomatic with hyperglycerolemia and hyperglyceroluria only.
Sequence similaritiesBelongs to the FGGY kinase family.
Cellular localizationMitochondrion outer membrane. Cytoplasm. In sperm and fetal tissues, the majority of the enzyme is bound to mitochondria, but in adult tissues, such as liver found in the cytoplasm.
- Information by UniProt
- ATP glycerol 3 phosphotransferase
- ATP:glycerol 3 phosphotransferase
- ATP:glycerol 3-phosphotransferase
KO cell lines
Our Abpromise guarantee covers the use of ab257966 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: 61 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:GK knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3:HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)
Lane 4:Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate (20 ug)
ab126599 was shown to specifically react with Glycerol kinase in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab267328 (knockout cell lysate ab257966) was used. Wild-type and Glycerol kinase knockout samples were subjected to SDS-PAGE. ab126599 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 500 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 insertion in exon4
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.
ab257966 has not yet been referenced specifically in any publications.