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AKT3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

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Images

Western blot - Human AKT3 knockout A549 cell line (AB286451), expandable thumbnail
  • Next Generation Sequencing - Human AKT3 knockout A549 cell line (AB286451), expandable thumbnail

Key facts

Cell type
A549
Species or organism
Human
Tissue
Lung
Form
Liquid
Knockout validation
Next Generation Sequencing

Alternative names

AKT, AKT3 kinase, AKT3_HUMAN, DKFZp434N0250, MPPH, PKB-GAMMA, PKBG, PRKBG, Protein kinase Akt-3, Protein kinase B gamma, RAC-PK-gamma, RAC-gamma, RAC-gamma serine/threonine-protein kinase, STK-2, Serine threonine protein kinase Akt 3, V akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma), V akt murine thymoma viral oncogene homolog 3 protein kinase B gamma, V-AKT murine thymoma viral oncogene homolog 3

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AKT3 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

Key facts

Cell type
A549
Form
Liquid
Disease
Carcinoma
Concentration
Loading...

Properties

Gene name
AKT3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing

Quality control

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

Cell culture

Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male

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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type A549 cell line (ab288558). 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 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

AKT3 also known as protein kinase B gamma plays an important role in various cellular processes. This serine/threonine kinase with a molecular mass of approximately 56 kDa functions as part of the AKT family which includes AKT1 and AKT2. AKT3 expresses highly in the brain tissues with significant expression also observed in the heart and kidneys. As an isoform of the AKT kinase it participates in signaling pathways essential for cell survival and metabolism.

Biological function summary

AKT3 participates in regulating cell proliferation and survival. It operates as part of the PI3K/AKT signaling complex a central hub for signaling growth factors. This pathway is critical for transmitting signals from the cell surface to the nucleus affecting transcription and cell cycle progression. AKT3 protein specifically contributes significant roles in neuronal development and function due to its high expression within the nervous system.

Pathways

AKT3 influences the PI3K/AKT and mTOR pathways. These pathways regulate cell growth metabolism and survival playing significant roles in normal cellular functions and cancer. AKT3 interacts closely with other signaling molecules including mTORC1 and mTORC2 complexes. While participating in these pathways AKT3 coordinates with the protein E17K which is pattern related with alterations leading to abnormal signaling and growth.

Associated diseases and disorders

AKT3 is implicated in cancer and neurological disorders. Aberrations in its signaling can lead to tumorigenesis as seen in glioblastomas. Abnormal AKT3 activity associates with epilepsy and other neurodevelopmental disorders emphasizing its impact on neuronal functions. Connecting AKT3 with disorders the protein E17K mutation in AKT3 dysregulates normal kinase activity contributing to these pathologies.

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.

2 product images

  • Western blot - Human AKT3 knockout A549 cell line (ab286451), expandable thumbnail

    Western blot - Human AKT3 knockout A549 cell line (ab286451)

    All lanes: Akt3 (E1Z3W) Rabbit mAb at 1/1000 dilution

    Lane 1: Wild-type A549 ab288558 at 20 µg

    Lane 2: AKT3 knockout A549 ab286451 at 20 µg

    Lane 3: THP-1 at 20 µg

    Lane 4: A549 Cytoplasmic at 20 µg

    Lane 5: Recombinant Human AKT3 protein at 0.2 µg

    Secondary

    All lanes: Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 54 kDa

    Observed band size: 60 kDa

  • Next Generation Sequencing - Human AKT3 knockout A549 cell line (ab286451), expandable thumbnail

    Next Generation Sequencing - Human AKT3 knockout A549 cell line (ab286451)

    68 bp deletion and 36 bp deletion after Arg325 (edit 1); 61 bp deletion after Val327 (edit 2)

Downloads

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'.

For licensing inquiries, please contact partnerships@abcam.com