Human CDC25C knockout HeLa cell line
- Advanced Validation
- What is this?
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CDC25C 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 insertion in exon 2 and 2 bp insertion in exon 2.
View Alternative Names
Cdc 25C, Cell division cycle 25 homolog C, Cell division cycle 25C, Cell division cycle 25C protein, Dual specificity phosphatase Cdc25C, M-phase inducer phosphatase 3, MPIP3_HUMAN, Mitosis inducer CDC25, PPP1R60, Phosphotyrosine phosphatase, protein phosphatase 1, regulatory subunit 60
- WB
Unknown
Western blot - Human CDC25C knockout HeLa cell line (AB265189)
Lanes 1- 2 : Merged signal (red and green). Green - ab32050 observed at 58 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32050 was shown to react with Cdc25C in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265189 (knockout cell lysate ab257387) was used. Wild-type HeLa and CDC25C 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. ab32050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 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-Cdc25C antibody [E303] (<a href='/en-us/products/primary-antibodies/cdc25c-antibody-e303-ab32050'>ab32050</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDC25C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDC25C knockout HeLa cell line (ab265189)
Predicted band size: 53 kDa
Observed band size: 58 kDa
false
- WB
Lab
Western blot - Human CDC25C knockout HeLa cell line (AB265189)
Lanes 1- 2 : Merged signal (red and green). Green - ab32444 observed at 58 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32444 was shown to react with Cdc25C in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265189 (knockout cell lysate ab257387) was used. Wild-type HeLa and CDC25C 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. ab32444 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 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-Cdc25C antibody [E302] (<a href='/en-us/products/primary-antibodies/cdc25c-antibody-e302-ab32444'>ab32444</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CDC25C knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CDC25C knockout HeLa cell line (ab265189)
Predicted band size: 53 kDa
Observed band size: 58 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CDC25C knockout HeLa cell line (AB265189)
Allele-1 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human CDC25C knockout HeLa cell line (AB265189)
Allele-2 : 2 bp insertion in exon 2.
- sELISA
Supplier Data
Sandwich ELISA - Human CDC25C knockout HeLa cell line (AB265189)
Human CDC25C concentration was interpolated from the CDC25C standard curve. Lysates from cell culture samples were serially diluted and assessed by the Human Cdc25C ELISA Kit (SimpleStep) (ab230944). Wild-type HeLa (ab255928), CDC25C knockout HeLa (ab265189) and Jurkat cells were assessed in duplicate (n=2). Data are represented as the mean and error bars represent standard deviation.
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
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
The proper function of Cdc25C involves facilitating proper cell cycle progression by coordinating with other cell cycle regulators. It participates in a complex network where it dephosphorylates and activates CDK1/cyclin B1 complexes promoting the mitotic entry. This function is important for maintaining genomic stability during cell division. Misregulation of Cdc25C can lead to cell cycle arrest or uncontrolled cell proliferation highlighting its essential role in cell cycle control mechanisms.
Pathways
Cdc25C fits into the cell cycle checkpoint pathways and is also a part of the DNA damage response pathways. It connects with the p53 and ATM/ATR signaling proteins while responding to DNA damage ensuring a temporary pause in cell cycle progression for repair mechanisms to act. Proper interaction with these pathways is essential for maintaining cellular integrity and preventing the proliferation of damaged cells with CDK1 and Wee1 kinase serving as major interacting proteins in these processes.
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
Complementary Products
![Western blot - Anti-Cdc25C antibody [E302] (AB32444)](https://content.abcam.com/products/images/cdc25c-antibody-e302-ab32444--western-blot-img95410.gif)
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