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AB257979

Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate

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GSK3A 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 deletion in exon1 and Insertion of the selection cassette in exon1.
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Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)
  • WB

Unknown

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)

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 : GSK3A knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab78664 was shown to specifically react with GSK3 alpha in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266742 (knockout cell lysate ab257979) was used. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab78664 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.

All lanes:

Western blot - Anti-GSK3 alpha antibody (<a href='/en-us/products/primary-antibodies/gsk3-alpha-antibody-ab78664'>ab78664</a>) at 1/1000 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:

GSK3A 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 GSK3A (GSK3 alpha) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gsk3a-gsk3-alpha-knockout-hek-293t-cell-line-ab266742'>ab266742</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) 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: 51 kDa

Observed band size: 51 kDa

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Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)
  • Sanger seq

Unknown

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)

Allele-2 : Insertion of the selection cassette in exon1

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)
  • Sanger seq

Unknown

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)

Allele-1 : 1 bp deletion in exon1

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)
  • WB

Lab

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (AB257979)

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 : GSK3A knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab40870 was shown to specifically react with GSK3 alpha in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266742 (knockout cell lysate ab257979) was used. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab40870 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.

All lanes:

Western blot - Anti-GSK3 alpha antibody [EP793Y] (<a href='/en-us/products/primary-antibodies/gsk3-alpha-antibody-ep793y-ab40870'>ab40870</a>) at 1/1000 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:

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell lysate (ab257979) at 20 µg

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) 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: 51 kDa

Observed band size: 51 kDa

false

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 deletion in exon1 and Insertion of the selection cassette in exon1.

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Complementary Products

Biochemicals

AB120962

SB 415286, GSK3 inhibitor

Chemical Structure - SB 415286, GSK3 inhibitor (AB120962)

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Proteins & Peptides

AB204149

Recombinant Human TSSK6 protein

SDS-PAGE - Recombinant Human TSSK6 protein (AB204149)

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Primary Antibodies

AB40870

Anti-GSK3 alpha antibody [EP793Y]

20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 alpha antibody [EP793Y] (AB40870)

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  • 1
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
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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?

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Properties and storage information

Gene name
GSK3A
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.

Glycogen synthase kinase 3 alpha (GSK3 alpha) is a serine/threonine protein kinase also known as GSK3A with a molecular mass of approximately 51 kDa. It plays an important role in the regulation of various cellular processes. GSK3 alpha is expressed in many tissues but has high levels in the brain heart and skeletal muscle. It participates in the phosphorylation of numerous substrates impacting their activity and stability.
Biological function summary

GSK3 alpha influences several cellular functions including cell division differentiation and apoptosis. It functions independently or as part of a protein complex often forming dimeric complexes with GSK3 beta. GSK3 alpha alters the activity of numerous transcription factors and regulates insulin and Wnt signaling pathways affecting glucose metabolism and cellular growth.

Pathways

GSK3 alpha is involved in the Wnt and insulin signaling pathways. In the Wnt pathway GSK3 alpha phosphorylates beta-catenin marking it for degradation and controlling gene expression. In the insulin pathway it modulates glycogen synthesis and affects glucose homeostasis. GSK3 alpha interacts closely with proteins like APC and glycogen synthase making it integral to these pathways.

GSK3 alpha links to neurodegenerative disorders and diabetes. Its dysfunction contributes to Alzheimer’s disease where abnormal phosphorylation of tau protein occurs. In the context of diabetes GSK3 alpha influences glucose regulation impacting insulin sensitivity and action. The interplay between GSK3 alpha and tau in Alzheimer's as well as insulin signaling proteins illustrates its important role in these disease pathways.
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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com