Human EIF2AK2 (PKR) knockout HeLa cell line
- Advanced Validation
- What is this?
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Human EIF2AK2 (PKR) knockout HeLa cell line available to order. Recommended control: Human wild-type HeLa cell line (ab255928).
View 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
- WB
Lab
Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
Lanes 1-5 : Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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-PKR antibody [YE350] (<a href='/en-us/products/primary-antibodies/pkr-antibody-ye350-ab32052'>ab32052</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (ab261824)
Lane 3:
Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 5:
K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 62 kDa
Observed band size: 70 kDa
false
- WB
Lab
Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
Lanes 1-5 : Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.
ab184257 Anti-PKR antibody [EPR19374] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab184257 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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-PKR antibody [EPR19374] (<a href='/en-us/products/primary-antibodies/pkr-antibody-epr19374-ab184257'>ab184257</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human EIF2AK2 (PKR) knockout HeLa cell line (ab261824)
Lane 3:
Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 5:
K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 62 kDa
Observed band size: 70 kDa
false
- Cell Culture
Lab
Cell Culture - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
Representative images EIF2AK2 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa 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 M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
Homozygous : 1 bp insertion in exon 1.
- Sanger seq
Lab
Sanger Sequencing - Human EIF2AK2 (PKR) knockout HeLa cell line (AB261824)
Sequencing chromatogram displaying sequence edit in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
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