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AB265069

Human FOXO3 (FOXO3A) knockout HEK-293T cell line

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FOXO3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/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

3 Images
Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)
  • WB

Lab

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109629 observed at 82 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab109629 was shown to react with FOXO3A in wild-type cells in Western blot with loss of signal observed in FOXO3 knockout cell lines. Wild-type and FOXO3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab109629 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h 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-293T cell lysate at 20 µg

Lane 2:

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-foxo3-foxo3a-knockout-hek-293t-cell-lysate-ab256922'>ab256922</a>) at 20 µg

Lane 2:

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (ab265069)

Lane 3:

Wild-type HEK-293 cell lysate at 20 µg

Lane 4:

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

Predicted band size: 71 kDa

Observed band size: 82 kDa

false

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)
  • WB

Lab

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)

Lanes 1 - 4 : Merged signal (red and green). Green - ab23683 observed at 82 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab23683 was shown to react with FOXO3A in wild-type cells in Western blot with loss of signal observed in FOXO3 knockout cell lines. Wild-type and FOXO3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab23683 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ° at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h 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-293T cell lysate at 20 µg

Lane 2:

Western blot - Human FOXO3 (FOXO3A) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-foxo3-foxo3a-knockout-hek-293t-cell-lysate-ab256922'>ab256922</a>) at 20 µg

Lane 3:

Wild-type HEK-293 cell lysate at 20 µg

Lane 4:

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

Predicted band size: 71 kDa

Observed band size: 82 kDa

false

Sanger Sequencing - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)
  • Sanger seq

Unknown

Sanger Sequencing - Human FOXO3 (FOXO3A) knockout HEK-293T cell line (AB265069)

Homozygous : 10 bp deletion in exon 1.

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, Homozygous: 10 bp deletion 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 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.

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What's included?

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Properties and storage information

Gene name
FOXO3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

FOXO3A also known as FOXO3 or 1e2 is a transcription factor with a molecular weight of approximately 71 kDa. It belongs to the Forkhead box O family of proteins. This protein plays an important role in the regulation of transcription in response to oxidative stress and growth stimuli. FOXO3A is expressed in a variety of tissues including liver muscle and brain where it contributes to a wide range of cellular processes related to stress resistance and metabolism. Scientists frequently study FOXO3A using techniques like FOXO3A ELISA and apply FOXO3A anticuerpos in research to understand its function more intricately.
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.

FOXO3A is linked to cancer and neurodegenerative diseases. In cancer alterations in FOXO3A activity or expression can affect tumor growth and resistance to oxidative stress often involving interactions with proteins like MDM2. Similarly in neurodegenerative disorders such as Alzheimer's disease FOXO3A modifies responses to oxidative damage potentially tied to proteins like Tau. Both instances highlight the role of FOXO3A as a versatile modulator that can influence the progression and management of various diseases.
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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

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Target data

See full target information FOXO3
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Secondary Antibodies

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Primary Antibodies

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Product promise

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