Human ATF3 knockout A549 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human ATF3 knockout A549 cell line (AB266955)
Lanes 1-2 : Merged signal (red and green). Green - ab207434 observed at 21 kDa. Red - loading control ab8245 observed at 37 kDa.
ab207434 Anti-ATF3 antibody [EPR19488] - ChIP Grade was shown to specifically react with ATF3 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266955 (knockout cell lysate ab257075) was used. Wild-type and ATF3 knockout samples were subjected to SDS-PAGE. ab207434 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 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-ATF3 antibody [EPR19488] - ChIP Grade (<a href='/en-us/products/primary-antibodies/atf3-antibody-epr19488-chip-grade-ab207434'>ab207434</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
ATF3 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human ATF3 knockout A549 cell line (ab266955)
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- WB
Lab
Western blot - Human ATF3 knockout A549 cell line (AB266955)
Lanes 1-2 : Merged signal (red and green). Green - ab254268 observed at 21 kDa. Red - loading control ab8245 observed at 37 kDa.
ab254268 Recombinant Anti-ATF3 antibody [EPR22610-19] was shown to specifically react with ATF3 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266955 (knockout cell lysate ab257075) was used. Wild-type and ATF3 knockout samples were subjected to SDS-PAGE. ab254268 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 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-ATF3 antibody [EPR22610-19] - ChIP Grade (<a href='/en-us/products/primary-antibodies/atf3-antibody-epr22610-19-chip-grade-ab254268'>ab254268</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
ATF3 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human ATF3 knockout A549 cell line (ab266955)
Predicted band size: 21 kDa
Observed band size: 21 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ATF3 knockout A549 cell line (AB266955)
Allele-2 : 4 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human ATF3 knockout A549 cell line (AB266955)
Allele-1 : 14 bp deletion in exon2
- Cell Culture
Unknown
Cell Culture - Human ATF3 knockout A549 cell line (AB266955)
Representative images of ATF3 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ATF3 knockout A549 cell line (AB266955)
Allele-3 : 1 bp deletion in exon 2.
Complementary Products
Primary Antibodies
AB207434
Anti-ATF3 antibody [EPR19488] - ChIP Grade
RabMAb|Advanced Validation|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-ATF3 antibody [EPR19488] - ChIP Grade (AB207434)](https://content.abcam.com/products/images/atf3-antibody-epr19488-chip-grade-ab207434--immunocytochemistry-immunofluorescence-img87225.jpg)
- 5
- 1
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.
- 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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
ATF3 acts as a regulator of cellular stress responses and can influence cell cycle arrest and apoptosis. It does not form a large complex but interacts with diverse partners to execute its functions. Studies often use HCT116 cells to investigate ATF3's role where it can serve as a marker of cellular stress and inflammation. The protein regulates genes involved in maintaining homeostasis in stressed cells.
Pathways
ATF3 is involved in stress response and apoptotic pathways. In the context of apoptosis ATF3 interacts with proteins like p53 and Jun which are important for inducing cell cycle arrest and programmed cell death. In stress response pathways ATF3 modifies transcriptional activity in reaction to various stress signals providing a checkpoint for damaged cells before repairing or entering programmed death.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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