Human ATF6 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human ATF6 knockout HeLa cell line (AB261800)
Lanes 1- 2 : Merged signal (red and green). Green - ab83504 observed at 95 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab83504 was shown to react with ATF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261800 (knockout cell lysate ab256841) was used. Wild-type HeLa and ATF6 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. ab83504 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at 1 μg/ml 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.
All lanes:
Western blot - Anti-ATF6 antibody (<a href='/en-us/products/primary-antibodies/atf6-antibody-ab83504'>ab83504</a>) at 1 µg/mL
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ATF6 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ATF6 knockout HeLa cell line (ab261800)
Predicted band size: 74 kDa
Observed band size: 95 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATF6 knockout HeLa cell line (AB261800)
Allele-2 : 17 bp deletion in exon 6.
- Sanger seq
Unknown
Sanger Sequencing - Human ATF6 knockout HeLa cell line (AB261800)
Allele-1 : 25 bp deletion in exon 6.
- Sanger seq
Unknown
Sanger Sequencing - Human ATF6 knockout HeLa cell line (AB261800)
Allele-3 : 4 bp deletion in exon 6.
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Cell Lines & Lysates
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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
ATF6 operates as part of the transcription regulation mechanisms responding to ER stress. The ATF6 protein is essential in managing the expression of chaperone genes and ER-associated degradation (ERAD) components thereby maintaining protein homeostasis. ATF6 itself does not operate within a traditional complex but its activation involves proteolytic cleavage which subsequently releases the active form to the nucleus where it influences gene expression to alleviate stress conditions.
Pathways
ATF6 is prominently involved in the unfolded protein response pathway which manages cell survival and stress adaptation. This pathway closely interacts with other proteins like IRE1 and PERK forming a network that modulates the transcription of UPR target genes. Additionally ATF6 contributes to checkpoint control pathways that stabilize cellular environment by regulating genes related to chaperone and protein folding.
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
Product promise
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