Human VCL (Vinculin) knockout A-431 cell line
- Advanced Validation
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VCL KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 16 bp deletion Frameshift = 99.96%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
CMD1W, CMH15, Epididymis luminal protein 114, HEL114, MV, MVCL, Metavinculin, OTTHUMP00000019861, OTTHUMP00000019862, VCL, VINC_HUMAN, Vinculin
- WB
Lab
Western blot - Human VCL (Vinculin) knockout A-431 cell line (AB261898)
Lanes 1 - 4 : Merged signal (red and green). Green - ab219649 observed at 124 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab219649 was shown to react with VCL in A431 wild-type cells in Western blot. Loss of signal was observed when VCL knockout sample was used. A431 wild-type and VCL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab219649 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Vinculin antibody [EPR20407] - Loading Control (<a href='/en-us/products/primary-antibodies/vinculin-antibody-epr20407-loading-control-ab219649'>ab219649</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human VCL (Vinculin) knockout A-431 cell line (ab261898)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Predicted band size: 124 kDa
Observed band size: 124 kDa
false
- WB
Lab
Western blot - Human VCL (Vinculin) knockout A-431 cell line (AB261898)
Western blot : Anti-VCL antibody [EPR8185] (ab129002) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab129002 was shown to bind specifically to VCL. A band was observed at 124 kDa in wild-type A431 cell lysates with no signal observed at this size in VCL knockout cell line. To generate this image, wild-type and VCL knockout A431 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°C. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (<a href='/en-us/products/primary-antibodies/vinculin-antibody-epr8185-loading-control-ab129002'>ab129002</a>) at 1/10000 dilution
Lane 1:
Wild-type A431 cell lysate
Lane 2:
Western blot - Human VCL (Vinculin) knockout A-431 cell line (ab261898) at 20 µg
Lane 2:
VCL knockout A431 cell lysate
Lane 3:
HeLa cell lysate
Lane 4:
Jurkat cell lysate
false
- WB
Lab
Western blot - Human VCL (Vinculin) knockout A-431 cell line (AB261898)
Lanes 1 - 4 : Merged signal (red and green). Green - ab207440 observed at 124 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab207440 was shown to react with VCL in A431 wild-type cells in Western blot. Loss of signal was observed when VCL knockout sample was used. A431 wild-type and VCL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab207440 and ab8245 (Mouse anti-GAPDH antibody [6C5]) 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) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Vinculin antibody [EPR19579] - Loading Control (<a href='/en-us/products/primary-antibodies/vinculin-antibody-epr19579-loading-control-ab207440'>ab207440</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human VCL (Vinculin) knockout A-431 cell line (ab261898)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Predicted band size: 124 kDa
Observed band size: 124 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human VCL (Vinculin) knockout A-431 cell line (AB261898)
8bp deletion after Phe126 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human VCL (Vinculin) knockout A-431 cell line (AB261898)
Knockout achieved by CRISPR/Cas9; X = 16 bp deletion; Frameshift = 99.96%
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB129002
Anti-Vinculin antibody [EPR8185] - Loading Control
20ul selling size|RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)](https://content.abcam.com/products/images/vinculin-antibody-epr8185-loading-control-ab129002--immunocytochemistry-immunofluorescence-img118915.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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