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AB261707

Human VCL (Vinculin) knockout A-431 cell lysate

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VCL KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 16 bp deletion Frameshift = 99.96%.

View Alternative Names

CMD1W, CMH15, Epididymis luminal protein 114, HEL114, MV, MVCL, Metavinculin, OTTHUMP00000019861, OTTHUMP00000019862, VCL, VINC_HUMAN, Vinculin

3 Images
Western blot - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)
  • WB

Lab

Western blot - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 2 : VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug

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 wild-type A-431 cells in Western blot Loss of signal was observed when VCL knockout cell line ab261898 (knockout cell lysate ab261707) was used. Wild-type A-431 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°C 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 (<a href='/en-us/products/cell-lines/human-vcl-vinculin-knockout-a-431-cell-line-ab261898'>ab261898</a>)

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

Western blot - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)
  • WB

Lab

Western blot - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 2 : VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug

Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug

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 wild-type A-431 cells in Western blot Loss of signal was observed when VCL knockout cell line ab261898 (knockout cell lysate ab261707) was used. Wild-type A-431 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°C 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 (<a href='/en-us/products/cell-lines/human-vcl-vinculin-knockout-a-431-cell-line-ab261898'>ab261898</a>)

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

Next Generation Sequencing - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)
  • NGS

Supplier Data

Next Generation Sequencing - Human VCL (Vinculin) knockout A-431 cell lysate (AB261707)

Knockout achieved by CRISPR/Cas9; X = 16 bp deletion; Frameshift = 99.96%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 16 bp deletion Frameshift = 99.96%

Disease

Epidermoid Carcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name
VCL
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

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