VCL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1
CMD1W, CMH15, Epididymis luminal protein 114, HEL114, MV, MVCL, Metavinculin, OTTHUMP00000019861, OTTHUMP00000019862, VCL, VINC_HUMAN, Vinculin
VCL KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
HeLa
Human
Cervix
Liquid
Sanger Sequencing, Western blot
Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1
Puromycin 1µg/mL
Adenocarcinoma
VCL
Knockout
CRISPR technology
Sanger Sequencing, Western blot
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
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.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Recommended control: Human wild-type HeLa cell line (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.
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Terms & Conditions.
Lanes 1-4: Merged signal (red and green). Green - Anti-Vinculin antibody [EPR19579] ab207440 observed at 124 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
Anti-Vinculin antibody [EPR19579] ab207440 Anti-Vinculin antibody [EPR19579] was shown to specifically react with Vinculin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265580 (knockout cell lysate Human VCL (Vinculin) knockout HeLa cell lysate ab257795) was used. Wild-type and Vinculin knockout samples were subjected to SDS-PAGE. Anti-Vinculin antibody [EPR19579] ab207440 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-Vinculin antibody [EPR19579] (Anti-Vinculin antibody [EPR19579] ab207440) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Vinculin knockout HeLa cell lysate at 20 µg
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 124 kDa
Observed band size: 124 kDa
Lanes 1-4: Merged signal (red and green). Green - Anti-Vinculin antibody [EPR19579] ab207440 observed at 124 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
Anti-Vinculin antibody [EPR19579] ab207440 Anti-Vinculin antibody [EPR19579] was shown to specifically react with Vinculin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265580 (knockout cell lysate Human VCL (Vinculin) knockout HeLa cell lysate ab257795) was used. Wild-type and Vinculin knockout samples were subjected to SDS-PAGE. Anti-Vinculin antibody [EPR19579] ab207440 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
Homozygous: 1 bp insertion in exon 1.
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