Human PSMD10 (Gankyrin) knockout HeLa cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell line (AB265680)
Lanes 1-4 : Merged signal (red and green). Green - ab188315 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab188315 Anti-Gankyrin antibody [EPR14456(2)] was shown to specifically react with Gankyrin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265680 (knockout cell lysate ab258146) was used. Wild-type and Gankyrin knockout samples were subjected to SDS-PAGE. ab188315 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-Gankyrin antibody [EPR14456(2)] (<a href='/en-us/products/primary-antibodies/gankyrin-antibody-epr144562-ab188315'>ab188315</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:
PSMD10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell line (ab265680)
Lane 3:
K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Lane 4:
HepG2 (Human liver hepatocellular carcinoma 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: 24 kDa
Observed band size: 24 kDa
false
- WB
Lab
Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell line (AB265680)
Lanes 1-4 : Merged signal (red and green). Green - ab182576 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab182576 Anti-Gankyrin antibody [EPR14455] was shown to specifically react with Gankyrin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265680 (knockout cell lysate ab258146) was used. Wild-type and Gankyrin knockout samples were subjected to SDS-PAGE. ab182576 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-Gankyrin antibody [EPR14455] (<a href='/en-us/products/primary-antibodies/gankyrin-antibody-epr14455-ab182576'>ab182576</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:
PSMD10 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PSMD10 (Gankyrin) knockout HeLa cell line (ab265680)
Lane 3:
K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole cell lysate at 20 µg
Lane 4:
HepG2 (Human liver hepatocellular carcinoma 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: 24 kDa
Observed band size: 24 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PSMD10 (Gankyrin) knockout HeLa cell line (AB265680)
Homozygous : 1 bp insertion in exon 1.
Complementary Products
Reactivity data
Product details
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
Gankyrin enhances the degradation of the retinoblastoma protein (pRb) as part of the 26S proteasome complex. This function disrupts normal cell cycle control promoting cell proliferation. Gankyrin binds directly to the cyclin-dependent kinase 4 (CDK4) and MDM2 which regulate pRb and p53 tumor suppressor pathways. Overexpression of Gankyrin contributes to cellular transformation and oncogenesis by regulating cell cycle progression and apoptosis.
Pathways
Gankyrin participates actively in p53 and ubiquitin-proteasome pathways. The protein facilitates ubiquitination and degradation of key regulators including p53 and pRb altering the stability of these proteins. Gankyrin interacts with proteins like MDM2 and HDAC1 influencing their roles in tumor development through these pathways. Its presence in these pathways contributes to unchecked cell growth by evading normal tumor suppressor mechanisms.
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 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