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AB257966

Human GK (Glycerol kinase) knockout HEK-293T cell lysate

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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.

View Alternative Names

ATP glycerol 3 phosphotransferase, ATP:glycerol 3-phosphotransferase, D930012N15Rik, GK, GK1, GKD, GLPK_HUMAN, Glycerokinase, Glycerol kinase, OTTHUMP00000023108, OTTHUMP00000023109, OTTHUMP00000215321

2 Images
Western blot - Human GK (Glycerol kinase) knockout HEK-293T cell lysate (AB257966)
  • WB

Lab

Western blot - Human GK (Glycerol kinase) knockout HEK-293T cell lysate (AB257966)

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.

All lanes:

Western blot - Anti-Glycerol kinase antibody [EPR6567] (<a href='/en-us/products/primary-antibodies/glycerol-kinase-antibody-epr6567-ab126599'>ab126599</a>) 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 (<a href='/en-us/products/cell-lines/human-gk-glycerol-kinase-knockout-hek-293t-cell-line-ab267328'>ab267328</a>)

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

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

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Sanger Sequencing - Human GK (Glycerol kinase) knockout HEK-293T cell lysate (AB257966)
  • Sanger seq

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Sanger Sequencing - Human GK (Glycerol kinase) knockout HEK-293T cell lysate (AB257966)

Homozygous : 1 bp insertion in exon4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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.

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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257966-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human GK knockout HEK293T cell lysate", "number":"AB257966-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257966-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
GK
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.
Biological function summary

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.

Pathways

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB126599

Anti-Glycerol kinase antibody [EPR6567]

RabMAb|Recombinant|KO Validated|Trial Size|Trial Size

Western blot - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

  • 5
  • 1
Cell Lines & Lysates

AB267328

Human GK (Glycerol kinase) knockout HEK-293T cell line

Advanced Validation

Western blot - Human GK (Glycerol kinase) knockout HEK-293T cell line (AB267328)

  • 0
  • 0

Product promise

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For full details, please see our Terms & Conditions

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