EIF2AK2 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 deletion in exon 5 and 1 bp insertion in exon 5.
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Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 1 bp insertion in exon 5
Alternative names
Double stranded RNA activated protein kinase;, E2AK2_HUMAN, EIF2AK1, EIF2AK2, Eukaryotic translation initiation factor 2-alpha kinase 2, HGNC:9437, Interferon inducible elF2 alpha kinase, Interferon-induced, double-stranded RNA-activated protein kinase, Interferon-inducible RNA-dependent protein kinase, MGC126524, P1/eIF-2A protein kinase, PPP1R83, PRKR, Protein kinase RNA-activated, Protein kinase interferon inducible double stranded RNA dependent, Protein phosphatase 1 regulatory subunit 83, Serine/threonine protein kinase TIK, Tyrosine protein kinase EIF2AK2, eIF-2A protein kinase 2, p68 kinase
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EIF2AK2 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 deletion in exon 5 and 1 bp insertion in exon 5.
Key facts
- Cell type
A549
- Species or organism
Human
- Tissue
Lung
- Form
Liquid
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 5 and 1 bp insertion in exon 5
- Antibiotic resistance
Puromycin 1µg/mL
- Disease
Carcinoma
- Concentration
Properties
- Gene name
EIF2AK2
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 1 US: 1
- Adherent/suspension
Adherent
- Gender
Male
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.
Storage
- Shipped at conditions
Dry Ice
- Appropriate long-term storage conditions
-196°C
Notes
Recommended control: Human wild-type A549 cell line (Human wild-type A549 cell line ab255450). 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.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human EIF2AK2 (PKR) knockout A549 cell line (ab266999)
Lanes 1-3: Merged signal (red and green). Green - Anti-PKR antibody [YE350] ab32052 observed at 70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-PKR antibody [YE350] ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266999 (knockout cell lysate Human EIF2AK2 (PKR) knockout A549 cell lysate ab256900) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. Anti-PKR antibody [YE350] ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-PKR antibody [YE350] (Anti-PKR antibody [YE350] ab32052) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: EIF2AK2 knockout A549 cell lysate at 20 µg
Lane 3: K-562 cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 62 kDa
Observed band size: 70 kDa
This data was developed using the same antibody clone in a different buffer formulation (Anti-PKR antibody [YE350] ab32052).
Lanes 1-3: Merged signal (red and green). Green - Anti-PKR antibody [YE350] ab32052 observed at 70 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-PKR antibody [YE350] ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266999 (knockout cell lysate Human EIF2AK2 (PKR) knockout A549 cell lysate ab256900) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. Anti-PKR antibody [YE350] ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
Sanger Sequencing - Human EIF2AK2 (PKR) knockout A549 cell line (ab266999)
Allele-1: 1 bp deletion in exon5
Sanger Sequencing - Human EIF2AK2 (PKR) knockout A549 cell line (ab266999)
Allele-2: 1 bp insertion in exon 5.
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