Human FOXO3 (FOXO3A) knockout HEK-293 cell line
- Advanced Validation
- What is this?
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FOXO3 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 15 bp deletion, 3 bp deletion. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
AF6q21, AF6q21 protein, DKFZp781A0677, FKHR2, FKHRL 1, FKHRL1P2, FOXO2, FOXO3_HUMAN, Forkhead (Drosophila) homolog (rhabdomyosarcoma) like 1, Forkhead Drosophila homolog of in rhabdomyosarcoma like 1, Forkhead box O3, Forkhead box O3A, Forkhead box protein O3, Forkhead box protein O3A, Forkhead homolog (rhabdomyosarcoma) like 1, Forkhead in rhabdomyosarcoma-like 1, MGC12739, MGC31925
- WB
Lab
Western blot - Human FOXO3 (FOXO3A) knockout HEK-293 cell line (AB260857)
Lanes 1 - 4 : Merged signal (red and green). Green - ab23683 observed at 71 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab23683 was shown to recognize FOXO3A in wild-type HEK-293 cells as signal was lost at the expected MW in FOXO3 knockout cell line ab260857 (knockout cell lysate ab261649). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and FOXO3 knockout samples were subjected to SDS-PAGE. ab23683 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-FOXO3A antibody (<a href='/en-us/products/primary-antibodies/foxo3a-antibody-ab23683'>ab23683</a>) at 1 µg/mL
Lane 1:
Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human FOXO3 (FOXO3A) knockout HEK-293 cell lysate (<a href='/en-us/products/cell-lysates/human-foxo3-foxo3a-knockout-hek-293-cell-lysate-ab261649'>ab261649</a>) at 20 µg
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Predicted band size: 71 kDa
Observed band size: 82 kDa
false
- WB
Lab
Western blot - Human FOXO3 (FOXO3A) knockout HEK-293 cell line (AB260857)
Lanes 1 - 4 : Merged signal (red and green). Green - ab109629 observed at 71 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109629 was shown to recognize FOXO3A in wild-type HEK-293 cells as signal was lost at the expected MW in FOXO3 knockout cell line ab260857 (knockout cell lysate ab261649). Additional cross-reactive bands were observed in the wild-type and knockout cell lysate. Wild-type and FOXO3 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab109629 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-FOXO3A antibody [EPR1950] (<a href='/en-us/products/primary-antibodies/foxo3a-antibody-epr1950-ab109629'>ab109629</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK 293 whole cell lysate at 20 µg
Lane 2:
FOXO3 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human FOXO3 (FOXO3A) knockout HEK-293 cell line (ab260857)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Predicted band size: 71 kDa
false
Reactivity data
Product details
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.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
FOXO3A acts as a regulator of genes involved in cell cycle arrest apoptosis and DNA repair. It is not part of a simple protein complex but works closely with multiple factors to influence cellular homeostasis. Significantly FOXO3A controls the expression of Bcl-2 a protein important for cell survival and the protein p27 which is important for cell cycle control. FOXO3A's activity modulates cellular longevity and impacts stress responses highlighting its abundant biological functions.
Pathways
FOXO3A integrates into the PI3K/AKT and the insulin signaling pathways both essential for cellular metabolism and proliferation control. In the PI3K/AKT pathway FOXO3A functionally links with proteins such as AKT and PTEN where its activity can be inhibited by AKT-mediated phosphorylation. In doing so it supports a balance between cell survival and apoptosis allowing the cell to adjust to growth conditions and external stressors effectively.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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