Human CDKN1C (p57 Kip2) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human CDKN1C (p57 Kip2) knockout HeLa cell line (AB280061)
False colour image of Western blot : Anti-p57 Kip2 antibody [EP2516] staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab119989 was shown to bind specifically to p57 Kip2. A band was observed at 50 kDa in wild-type HeLa cell lysates with no signal observed at this size in CDKN1C knockout cell line ab280061 (knockout cell lysate ab280120). To generate this image wild-type and CDKN1C knockout HeLa cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) at 1/20000 dilution.
All lanes:
Western blot - Anti-p57 Kip2 antibody [EP2516] (<a href='/en-us/products/primary-antibodies/p57-kip2-antibody-ep2516-ab119989'>ab119989</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277359 cell lysate at 20 µg
Lane 2:
Wild-type HeLa Treated Dexamethasone (50 nM, 16 h) ab287335 cell lysate at 20 µg
Lane 2:
Western blot - Human CDKN1C (p57 Kip2) knockout HeLa cell line (ab280061)
Lane 3:
CDKN1C knockout HeLa Vehicle Control Dexamethasone (0 nM, 16 h) ab277299 cell lysate at 20 µg
Lane 4:
CDKN1C knockout HeLa Treated Dexamethasone (50 nM, 16 h) ab281877 cell lysate at 20 µg
Lane 5:
MCF7 cell lysate at 20 µg
Lane 6:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 32 kDa
Observed band size: 50 kDa
false
- Sanger seq
Lab
Sanger Sequencing - Human CDKN1C (p57 Kip2) knockout HeLa cell line (AB280061)
217 bp deletion in exon 1
Complementary Products
Primary Antibodies
AB133531
Anti-p57 Kip2 antibody [EP2718(2)]
BOND RX™ Validated|RabMAb|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p57 Kip2 antibody [EP2718(2)] (AB133531)](https://content.abcam.com/products/images/p57-kip2-antibody-ep27182-ab133531--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img439025.jpg)
- 5
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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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
P57^Kip2 acts as a regulator of cell cycle progression. It binds directly to cyclin-CDK complexes stopping cell cycle transition from G1 to S phase. This action prevents unregulated cell division and contributes to cellular differentiation. p57^Kip2 forms part of larger protein complexes further influencing various cellular processes. Its activity is important for embryonic development where it controls cell proliferation rates.
Pathways
P57^Kip2 integrates into pathways controlling cell cycle checkpoints and apoptosis. It interacts with proteins like cyclin D and E and the retinoblastoma protein (Rb) within the cell cycle regulation pathway. Additionally its presence in growth signaling pathways intersects with TGF-beta signaling. These pathways hold significant influence over development and cellular responses to DNA damage.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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