AB266742

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

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GSK3A KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

5 Images

Lab

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Lanes 1-4 : Merged signal (red and green). Green - ab40870 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.

ab40870 Anti-GSK3 alpha antibody [EP793Y] 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 4° 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 (<a href='/en-us/products/cell-lysates/human-gsk3a-gsk3-alpha-knockout-hek-293t-cell-lysate-ab257979'>ab257979</a>) 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

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Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Lanes 1-4 : Merged signal (red and green). Green - ab78664 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.

ab78664 Anti-GSK3 alpha antibody 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 4° 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 (ab266742)

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:

Predicted band size: 51 kDa

Observed band size: 51 kDa

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Cell Culture - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Cell Culture

Lab

Cell Culture - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Representative images GSK3A knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Sanger seq

Unknown

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Allele-2 : Insertion of the selection cassette in exon 1.

Sanger seq

Unknown

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Allele-1 : 1 bp deletion in exon1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

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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

We will provide viable cells that proliferate on revival.

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

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO₂. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10⁴ cells/cm². Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines

All seeding densities should be based on cell counts gained by established methods.
A guide seeding density of 2x10⁴ cells/cm² is recommended.
Cells should be passaged when they have achieved 80-90% confluence.

Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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

Visit the General protocols
Visit the Troubleshooting

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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

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

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Cell Culture - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Complementary Products

Primary Antibodies

Primary Antibodies

Biochemicals

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Handling procedures

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise