Human EIF2AK3 knockout A549 cell line
- Advanced Validation
- What is this?
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EIF2AK3 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
DKFZp781H1925, E2AK3_HUMAN, EC 2.7.11.1, Eif2ak3, Eukaryotic translation initiation factor 2-alpha kinase 3, HRI, Heme regulated EIF2 alpha kinase, HsPEK, PEK, PRKR-like endoplasmic reticulum kinase, Pancreatic eIF2-alpha kinase, WRS
- WB
Lab
Western blot - Human EIF2AK3 knockout A549 cell line (AB288905)
Western blot : Anti-TET2 antibody (ab124297) staining at 1/250 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab124297 was shown to bind specifically to TET2. 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-PERK antibody [EPR19876-294] (<a href='/en-us/products/primary-antibodies/perk-antibody-epr19876-294-ab229912'>ab229912</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
EIF2AK3 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human EIF2AK3 knockout A549 cell line (ab288905)
Lane 3:
MCF7 cell lysate at 20 µg
Lane 4:
Human eye cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 145 kDa
false
- WB
Lab
Western blot - Human EIF2AK3 knockout A549 cell line (AB288905)
Western blot : Anti-EIF2AK3 antibody [EPR19876-294] (ab229912) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab229912 was shown to bind specifically to EIF2AK3. A band was observed at 145 kDa in wild-type A549 cell lysates with no signal observed at this size in EIF2AK3 knockout cell line. To generate this image, wild-type and EIF2AK3 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-PERK antibody (<a href='/en-us/products/primary-antibodies/perk-antibody-ab65142'>ab65142</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
EIF2AK3 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human EIF2AK3 knockout A549 cell line (ab288905)
Lane 3:
MCF7 cell lysate at 20 µg
Lane 4:
Human eye cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 145 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human EIF2AK3 knockout A549 cell line (AB288905)
24 bp deletion after His 262, and 52 bp deletion after Ala 314 (allele 1); 1 bp deletion after Pro 269, 52 deletion downstream (allele 2); 181 bp insertion after Asp 270, 180 bp deletion downstream (allele 3)
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.
Recommended control: Human wild-type A549 cell line (ab288558). 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 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
PERK plays an important role in maintaining cellular homeostasis during stress conditions. It forms part of a larger complex that regulates the adaptive response to ER stress. Upon activation PERK arrests general protein translation while allowing for selective translation of stress response proteins. This mechanism prevents the accumulation of unfolded proteins and assists in the survival of cells under adverse conditions. PERK also contributes to the regulation of oxidative stress and cell apoptosis adding further layers to its biological significance.
Pathways
PERK's activity intersects with the integrated stress response and UPR pathways. It shares a close functional relationship with proteins like ATF4 which is synthesized upon PERK activation and is important for the transcription of genes related to stress adaptation. The UPR pathway coordinates with the IRE1 and ATF6 pathways forming an important network for managing ER stress. These pathways collectively ensure cellular resilience adjusting metabolic demands and promoting cellular recovery.
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
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Advanced Validation
- 0
- 0
Primary Antibodies
AB229912
Anti-PERK antibody [EPR19876-294]
20ul selling size|RabMAb|Recombinant|KO Validated
![Western blot - Anti-PERK antibody [EPR19876-294] (AB229912)](https://content.abcam.com/products/images/perk-antibody-epr19876-294-ab229912--western-blot-img444980.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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