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AB261823

Human FOXA1 knockout HeLa cell line

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FOXA1 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 insertion in exon 2 and 4 bp insertion in exon 2.

View Alternative Names

FOXA1_HUMAN, Forkhead box protein A1, HNF-3-alpha, HNF-3A, Hepatocyte nuclear factor 3-alpha, MGC33105, TCF-3A, Transcription factor 3A, forkhead box A1

4 Images
Western blot - Human FOXA1 knockout HeLa cell line (AB261823)
  • WB

Lab

Western blot - Human FOXA1 knockout HeLa cell line (AB261823)

Lanes 1- 2 : Merged signal (red and green). Green - ab170933 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab170933 was shown to react with FOXA1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261823 (knockout cell lysate ab256920) was used. Wild-type HeLa and FOXA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170933 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Hela cells are predicted to express FOXA1, not FOXA2 and FOXA3

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (<a href='/en-us/products/primary-antibodies/foxa1-antibody-epr10881-ab170933'>ab170933</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FOXA1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOXA1 knockout HeLa cell line (ab261823)

Predicted band size: 49 kDa

Observed band size: 52 kDa

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Western blot - Human FOXA1 knockout HeLa cell line (AB261823)
  • WB

Lab

Western blot - Human FOXA1 knockout HeLa cell line (AB261823)

Lanes 1- 2 : Merged signal (red and green). Green - ab55178 observed at 49 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

ab55178 was shown to react with FOXA1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261823 (knockout cell lysate ab256920) was used. Wild-type HeLa and FOXA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab55178 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-FOXA1 antibody [1B1] (<a href='/en-us/products/primary-antibodies/foxa1-antibody-1b1-ab55178'>ab55178</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FOXA1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOXA1 knockout HeLa cell line (ab261823)

Predicted band size: 49 kDa

Observed band size: 49 kDa

false

Sanger Sequencing - Human FOXA1 knockout HeLa cell line (AB261823)
  • Sanger seq

Unknown

Sanger Sequencing - Human FOXA1 knockout HeLa cell line (AB261823)

Allele-2 : 4 bp insertion in exon 2.

Sanger Sequencing - Human FOXA1 knockout HeLa cell line (AB261823)
  • Sanger seq

Unknown

Sanger Sequencing - Human FOXA1 knockout HeLa cell line (AB261823)

Allele-1 : 1 bp insertion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 4 bp insertion in exon 2

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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.

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What's included?

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Properties and storage information

Gene name
FOXA1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

FOXA1 also known as Forkhead Box A1 is a transcription factor with a molecular weight of about 49 kDa. It plays a role by binding to specific DNA sequences influencing the expression of genes involved in various cellular processes. This protein is expressed in tissues like liver lungs and prostate. Researchers also refer to FOXA1 as HNF-3α emphasizing its importance in transcription regulation.
Biological function summary

FOXA1 influences the transcriptional control of genes essential for organ development and metabolic regulation. It acts as a pioneer factor opening chromatin structure to enable access for other transcription factors. FOXA1 integrates into complexes that involve histone modification affecting the chromatin state and transcription of target genes. This function is necessary for the proper differentiation and function of several tissues.

Pathways

FOXA1 participates significantly in the androgen receptor signaling pathway and the estrogen receptor signaling pathway. It interacts closely with androgen receptor facilitating the expression of genes that influence cell growth and proliferation. The involvement of FOXA1 in these pathways highlights its critical role in pathways that regulate hormone-responsive cancers and metabolic processes.

FOXA1 shows a strong association with cancers such as prostate cancer and breast cancer. FOXA1's interaction with the androgen receptor links it to prostate cancer progression while its involvement with the estrogen receptor correlates with breast cancer development. Aberrations in its expression or function can disrupt these pathways emphasizing the importance of FOXA1 in cancer biology and its potential as a therapeutic target.
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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 FOXA1
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Western blot - Human TUBB3 (beta III Tubulin) knockout HeLa cell line (AB255358)

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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

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