GK KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
ATP glycerol 3 phosphotransferase, ATP:glycerol 3-phosphotransferase, D930012N15Rik, GK, GK1, GKD, GLPK_HUMAN, Glycerokinase, Glycerol kinase, OTTHUMP00000023108, OTTHUMP00000023109, OTTHUMP00000215321
GK KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
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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. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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Glycerol kinase also known as GK or Gyk acts as a catalyst in the phosphorylation of glycerol to glycerol-3-phosphate. This enzymatic activity is important for the utilization of glycerol in metabolic processes. Glycerol kinase is expressed in tissues such as liver kidney and adipose tissue. Its molecular weight varies depending on the species but in humans it is around 60 kDa when analyzed using methods like glycerol SDS PAGE. In human cells like HEK293T reliable detection of glycerol kinase is possible due to its considerable expression.
The main role of glycerol kinase relates to its function in lipid metabolism and energy homeostasis. It exists as a part of a biochemical pathway but does not typically form a complex with other proteins. Glycerol kinase enables cells to utilize glycerol as an energy source by converting it into glycerol-3-phosphate which feeds into further metabolic pathways. This conversion is essential for the balance of energy supply and the production of triglycerides and phospholipids.
Metabolism and energy production heavily rely on the function of glycerol kinase. This enzyme acts within the glycerolipid metabolism and gluconeogenesis pathways. In these pathways it is connected to other enzymes like glycerol-3-phosphate dehydrogenase which further processes the glycerol-3-phosphate produced by glycerol kinase. The enzyme's availability affects the rate of these pathways and in turn influences lipid and glucose metabolism in the body.
Glycerol kinase deficiency leads to complex metabolic syndromes such as hyperglycerolemia and congenital adrenal hyperplasia. Hyperglycerolemia results from excess glycerol in the blood due to impaired conversion by the enzyme. In congenital adrenal hyperplasia alterations in metabolic function relate to glycerol kinase’s impact on stress response and energy balance. Disorders related to glycerol kinase sometimes involve interactions with other proteins and enzymes involved in broader metabolic processes affecting overall metabolism and physiology.
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Anti-Glycerol kinase antibody [EPR6567] ab126599 was shown to specifically react with Glycerol kinase in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human GK (Glycerol kinase) knockout HEK-293T cell line ab267328 (knockout cell lysate ab257966) was used. Wild-type and Glycerol kinase knockout samples were subjected to SDS-PAGE. Anti-Glycerol kinase antibody [EPR6567] ab126599 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-Glycerol kinase antibody [EPR6567] (Anti-Glycerol kinase antibody [EPR6567] ab126599) at 1/500 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: GK knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: Western blot - Human GK (Glycerol kinase) knockout HEK-293T cell line (Human GK (Glycerol kinase) knockout HEK-293T cell line ab267328)
Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
Homozygous: 1 bp insertion in exon4
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