Human RBL1 (p107) knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human RBL1 (p107) knockout HeLa cell line (AB264706)
False colour image of Western blot : Anti-p107 antibody staining at 1/400 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab245514 was shown to bind specifically to p107. A band was observed at 121 kDa in wild-type HeLa cell lysates with no signal observed at this size in RBL1 knockout cell line ab264706 (knockout cell lysate ab258630). To generate this image wild-type and RBL1 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 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-p107 antibody (<a href='/en-us/products/primary-antibodies/p107-antibody-ab245514'>ab245514</a>) at 1/400 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RBL1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RBL1 (p107) knockout HeLa cell line (ab264706)
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 121 kDa
Observed band size: 121 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RBL1 (p107) knockout HeLa cell line (AB264706)
Homozygous : 1 bp insertion in exon 9.
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
Retinoblastoma-like protein 1 functions in the regulation of the cell cycle acting as an important checkpoint controller between the G1 and S phases. This protein forms a complex with E2F transcription factors which are important in controlling genes involved in cell cycle progression. The p107-E2F interaction represses transcription and inhibits the cell from proceeding to the S phase thereby ensuring proper cellular proliferation control. Moreover p107 can bind to cyclin-dependent kinases adding another layer of regulation.
Pathways
P107 plays essential roles in the RB-E2F pathway and the DNA damage response pathway. Within these pathways p107 interacts with other key proteins like p130 and pRB which together form the pocket protein family. The RB-E2F pathway controls cell cycle entry and exit while the DNA damage response pathway ensures genomic integrity. p107 acts as a mediator to pause the cell cycle during DNA damage repair collaborating with these related proteins to maintain cellular homeostasis.
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'.
For licensing inquiries, please contact partnerships@abcam.com