Human NFE2L2 knockout A549 cell line[21]
- Advanced Validation
- What is this?
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(0 Publication)
![Western blot - Human NFE2L2 knockout A549 cell line[21] (AB285359)](https://content.abcam.com/products/images/human-nfe2l2-knockout-a549-cell-line21-ab285359--western-blot-img427758.jpg/_jcr_content/renditions/webp-768.webp)
- WB
Supplier Data
Western blot - Human NFE2L2 knockout A549 cell line[21] (AB285359)
Western blot : Anti-NFE2L2 antibody (ab137550) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137550 was shown to bind specifically to NFE2L2. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in NFE2L2 knockout cell line ab285359 (knockout cell lysate ab289682). To generate this image, wild-type and NFE2L2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
false
![Western blot - Human NFE2L2 knockout A549 cell line[21] (AB285359)](https://content.abcam.com/products/images/human-nfe2l2-knockout-a549-cell-line21-ab285359--western-blot-img416794.jpg/_jcr_content/renditions/webp-768.webp)
- WB
Supplier Data
Western blot - Human NFE2L2 knockout A549 cell line[21] (AB285359)
Western blot : Anti-NFE2L2 antibody (ab137550) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137550 was shown to bind specifically to NFE2L2. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in NFE2L2 knockout cell line ab285359 (knockout cell lysate ab289682). To generate this image, wild-type and NFE2L2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Nrf2 antibody (<a href='/en-us/products/primary-antibodies/nrf2-antibody-ab137550'>ab137550</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
NFE2L2 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human NFE2L2 knockout A549 cell line[21] (ab285359)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
Huh7 cell lysate at 20 µg
Predicted band size: 68 kDa
Observed band size: 85 kDa
false
![Sanger Sequencing - Human NFE2L2 knockout A549 cell line[21] (AB285359)](https://content.abcam.com/products/images/human-nfe2l2-knockout-a549-cell-line21-ab285359--sanger-sequencing-img427886.jpg/_jcr_content/renditions/webp-768.webp)
- Sanger seq
Supplier Data
Sanger Sequencing - Human NFE2L2 knockout A549 cell line[21] (AB285359)
This cell line contains three unique indels : -101bp, -103bp, and -1bp deletion in exon 4 of the NFE2L2 gene.
Complementary Products
![Western blot - Anti-Nrf2 antibody [EP1808Y] (AB62352)](https://content.abcam.com/products/images/nrf2-antibody-ep1808y-ab62352--western-blot-img417623.jpg)
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
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., Cells should be passaged when they have achieved 80-90% confluence (approx 8x104-1x105 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
Nrf2 acts as an important regulator of the cellular antioxidant response. It works in conjunction with its partner protein Keap1 forming a complex that controls its stability and degradation. Under normal conditions Keap1 keeps Nrf2 in the cytoplasm where it is targeted for ubiquitination and degradation. Once activated by oxidative stress or electrophiles Nrf2 dissociates from Keap1 thereby avoiding degradation and relocates to the nucleus to activate the transcription of ARE-dependent genes. This activity boosts the cellular response to oxidative stress by inducing genes involved in detoxification and cellular defense.
Pathways
Nrf2 plays a significant role in the oxidative stress response and detoxification pathways. Nrf2 activation is linked closely to the PI3K/Akt signaling pathway which influences cell survival growth and metabolism. This pathway also interacts with other important proteins like GSK-3β which can modulate Nrf2 activity and stability. Through these pathways Nrf2 orchestrates a defense mechanism against reactive oxygen species (ROS) by boosting the expression of antioxidant enzymes and detoxifying proteins.
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'.
For licensing inquiries, please contact partnerships@abcam.com