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GSK3A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. 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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Images

Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742), expandable thumbnail
  • Western blot - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742), expandable thumbnail
  • Cell Culture - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742), expandable thumbnail
  • Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742), expandable thumbnail
  • Sanger Sequencing - Human GSK3A (GSK3 alpha) knockout HEK-293T cell line (AB266742), expandable thumbnail

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

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

Alternative names

Recommended products

GSK3A KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

Key facts

Cell type

HEK-293T

Form

Liquid

Mutation description

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

Concentration
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Properties

Gene name

GSK3A

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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.

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (Human wild-type HEK-293T cell line ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

Supplementary info

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

Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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

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