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Human AKT1 mutation MCF7 cell line available to order.

Recommended control: Human wild-type MCF7 cell line (ab277916).

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Images

Sanger Sequencing - Human AKT1 mutation MCF7 cell line (AB308495), expandable thumbnail

Key facts

Cell type
MCF7
Species or organism
Human
Tissue
Breast
Form
Liquid
Knockout validation
Sanger Sequencing
Mutation description
Homozygote. pAKT1(Ser473) deletion

Alternative names

AKT, AKT1_HUMAN, C AKT, MGC99656, PKB, PKB-ALPHA, PRKBA, Protein Kinase B Alpha, Protein kinase B, Proto-oncogene c-Akt, RAC, RAC Serine/Threonine Protein Kinase, RAC-ALPHA, RAC-PK-alpha, RAC-alpha serine/threonine-protein kinase, Rac protein kinase alpha, V-AKT murine thymoma viral oncogene homolog 1

Human AKT1 mutation MCF7 cell line available to order.

Recommended control: Human wild-type MCF7 cell line (ab277916).

Key facts

Cell type
MCF7
Form
Liquid
Mutation description
Homozygote. pAKT1(Ser473) deletion
Disease
Adenocarcinoma
Concentration
Loading...

Properties

Gene name
AKT1
Gene editing type
Mutation
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing

Cell culture

Biosafety level
EU: 1 US: 1
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
  • Slow to trypsinise.
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 5-7x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
EMEM + 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 duration
A few days
Appropriate short-term storage conditions
-80°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type MCF7 cell line (ab277916). 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.

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

Culture medium: EMEM + 10% FBS

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. 
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:

• Slow to trypsinise
• All seeding densities should be based on cell counts gained by established methods.
• A guide seeding density of 5-7x104 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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

AKT1 also known as Protein Kinase B (PKB) is a serine/threonine kinase involved in various cellular processes. AKT1 plays an important role in mediating signals for cell survival growth and metabolism. This protein has a molecular weight of approximately 56 kDa. AKT1 is ubiquitously expressed in many tissues including the brain heart and lungs showing its importance in multiple physiological contexts. Phosphorylation of AKT1 at serine 473 denoted as p-AKT S473 is an important modification that regulates its activity.

Biological function summary

AKT1 regulates a broad spectrum of cellular functions through signaling pathways. It is an important player in the phosphatidylinositol 3-kinase (PI3K) and AKT pathway forming part of a complex involved in promoting cell survival and growth. AKT1 interacts closely with other proteins such as mTOR influencing cellular metabolism and autophagy. The phosphorylation state of AKT1 is critical for its activity with modifications like p-AKT S473 impacting its interaction with cellular substrates.

Pathways

AKT1 participates in the PI3K/AKT/mTOR signaling cascade instrumental in cell proliferation and metabolism. AKT1 integrates signals from insulin and growth factors modulating pathways that control cell growth and glucose uptake. This pathway involves proteins like mTOR and phospho-AKT which coordinate cellular responses to external stimuli. AKT1’s phosphorylation at serine 473 and the involvement of AKT1 E17K a gain-of-function mutation influence these pathways significantly.

Associated diseases and disorders

AKT1 has associations with cancer and metabolic disorders. Dysregulation of the PI3K/AKT/mTOR pathway involving AKT1 and mTOR often results in oncogenic transformation and uncontrolled cellular proliferation. In cancer such as breast cancer AKT1 mutations including AKT1 E17K are implicated altering cell signals for growth. Additionally AKT1 is connected to metabolic disorders such as Type 2 diabetes where it affects insulin signaling and glucose metabolism highlighting its interaction with metabolic proteins like mTOR and phospho-AKT.

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1 product image

  • Sanger Sequencing - Human AKT1 mutation MCF7 cell line (ab308495), expandable thumbnail

    Sanger Sequencing - Human AKT1 mutation MCF7 cell line (ab308495)

    Homozygote. pAKT1(Ser473) deletion, the PAM mutation is in Blue

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Product protocols

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

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