Human PDCD4 knockout HeLa cell line
- Advanced Validation
- What is this?
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PDCD4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 7.
View Alternative Names
DUG, Death up-regulated gene protein, H731, MA 3, MGC33046, MGC33047, Neoplastic transformation inhibitor, Neoplastic transformation inhibitor protein, Nuclear antigen H731, Nuclear antigen H731 like protein, Nuclear antigen H731-like, PDCD4_HUMAN, Programmed cell death 4, Programmed cell death protein 4, Protein 197/15a, Protein MA-3, Tis, Topoisomerase-inhibitor suppressed protein, programmed cell death 4 (neoplastic transformation inhibitor)
- WB
Lab
Western blot - Human PDCD4 knockout HeLa cell line (AB261833)
Lanes 1- 2 : Merged signal (red and green). Green - ab80590 observed at 51 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab80590 was shown to react with PDCD4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261833 (knockout cell lysate ab257278) was used. Wild-type HeLa and PDCD4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab80590 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 20000 dilution and a 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-PDCD4 antibody [EPR3431] (<a href='/en-us/products/primary-antibodies/pdcd4-antibody-epr3431-ab80590'>ab80590</a>) at 1/20000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
PDCD4 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human PDCD4 knockout HeLa cell line (ab261833)
Predicted band size: 51 kDa
Observed band size: 51 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PDCD4 knockout HeLa cell line (AB261833)
Homozygous : 1 bp insertion in exon 7.
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
PDCD4 inhibits translation by binding to the eIF4A RNA helicase blocking the translation initiation complex. This action stabilizes mRNA and reduces protein synthesis impacting processes such as cell growth and proliferation. Although PDCD4 functions mainly as a monomer it can also interact with other proteins to form complexes that enhance its regulatory activity. More specifically its role in controlling AP-1 transcription factor activity steps into the spotlight in influencing cellular responses.
Pathways
PDCD4 primarily interacts with signaling mechanisms related to apoptosis and inflammation. The protein is integral in the mTOR signaling pathway which governs cell growth in response to nutrient availability. PDCD4 also impacts the PI3K/AKT pathway linking its activity to cellular survival and metabolism. Through these pathways PDCD4 associates with proteins such as p53 and NF-kB connecting it to broader cellular and molecular networks involved in maintaining cellular integrity.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Primary Antibodies
AB80590
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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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