Human YES1 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human YES1 knockout HeLa cell line (AB265202)
Lanes 1- 4 : Merged signal (red and green). Green - ab109265 observed at 60 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109265 was shown to react with Yes1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab265202 (CRISPR/Cas9 edited cell lysate ab258290) lane below 60kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and YES1 CRISPR/Cas9 edited 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. ab109265 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-Yes1 antibody [EPR3173] (<a href='/en-us/products/primary-antibodies/yes1-antibody-epr3173-ab109265'>ab109265</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
YES1 CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human YES1 knockout HeLa cell line (ab265202)
Lane 3:
SW 480 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 60 kDa
Observed band size: 60 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human YES1 knockout HeLa cell line (AB265202)
Allele-1 : 31 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human YES1 knockout HeLa cell line (AB265202)
Allele-3 : 8 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human YES1 knockout HeLa cell line (AB265202)
Allele-2 : 14 bp deletion in exon 2.
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Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
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 Yes1 protein participates in cellular processes such as cell growth survival and differentiation. It contributes to signaling cascades that regulate these functions often working as part of protein complexes. Its ability to interact with various signaling molecules positions it as an important player in the control of cell behavior particularly in response to extracellular stimuli.
Pathways
The Yes1 protein integrates into key signaling routes like the epidermal growth factor receptor (EGFR) signaling pathway and the integrin signaling pathway. Within these pathways Yes1 interacts with proteins like c-Src and Fyn which are other members of the Src family. These pathways are vital in mediating cellular communication for growth and attachment highlighting Yes1's role in transmitting signals that prompt cellular responses.
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