Human CYR61 (CCN1) knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human CYR61 (CCN1) knockout HeLa cell line (AB265288)
Lanes 1-4 : Merged signal (red and green). Green - ab230947 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa.
ab230947 Anti-CYR61/CCN1 antibody [EPR20681] was shown to specifically react with CYR61/CCN1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265288 (knockout cell lysate ab257406) was used. Wild-type and CYR61/CCN1 knockout samples were subjected to SDS-PAGE. ab230947 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 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-CYR61/CCN1 antibody [EPR20681] (<a href='/en-us/products/primary-antibodies/cyr61-ccn1-antibody-epr20681-ab230947'>ab230947</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CYR61 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CYR61 (CCN1) knockout HeLa cell line (ab265288)
Lane 3:
MDA-MB-231 cell lysate at 20 µg
Lane 4:
MCF7 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: 42 kDa
Observed band size: 47 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell line (AB265288)
Allele-1 : 1 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell line (AB265288)
Allele-2 : Insertion of the selection cassette 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
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 CYR61/CCN1 protein acts as a signaling molecule that influences cell proliferation migration and adhesion. It interacts with integrins and heparan sulfate proteoglycans on the cell surface facilitating cellular communication that is essential for structural and functional tissue integrity. Additionally CYR61 is a part of multiprotein complexes impacting angiogenesis and inflammation. These activities place CYR61/CCN1 at a pivotal point in wound healing and vascularization processes.
Pathways
The CYR61/CCN1 protein participates in key signaling pathways such as the MAPK/ERK and Wnt pathways. These pathways modulate gene expression related to cell growth and differentiation. CYR61 collaborates with other proteins including VEGF and FGF to regulate cardiovascular development and morphogenesis. The interaction with these pathways highlights the protein's integral role in cellular homeostasis and response to physiological changes.
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