Human VIM (Vimentin) knockout HeLa cell lysate

3 이미지

• WB

Unknown

Western blot - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

This image was generated using a previous batch manufactured using the hybridoma production method.

Lane 1 : U20S cell lysate (20 μg)

Lane 2 : Hu tonsil cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : VIM knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab16700 observed at 53 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 100 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.

Lanes 1 - 4:

Western blot - Anti-Vimentin antibody [SP20], prediluted (<a href='/ko/products/primary-antibodies/vimentin-antibody-sp20-prediluted-ab27608'>ab27608</a>) at 1/100 dilution

Lanes 1 - 4:

Western blot - Anti-Vimentin antibody [SP20] (<a href='/ko/products/primary-antibodies/vimentin-antibody-sp20-ab16700'>ab16700</a>) at 1/100 dilution

Lane 1:

U20S cell lysate at 20 µg

Lane 2:

Human tonsil cell lysate at 20 µg

Lane 2:

Western blot - Human VIM (Vimentin) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-vim-vimentin-knockout-hela-cell-line-ab255446'>ab255446</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

VIM knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 37 kDa,53 kDa

false

• WB

Lab

Western blot - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

Lane 1 : U20S cell lysate (20 μg)

Lane 2 : Hu tonsil cell lysate (20 μg)

Lane 3 : Wild-type HeLa cell lysate (20 μg)

Lane 4 : VIM knockout HeLa cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab8069 observed at 53 kDa. Red - loading control, ab181602 observed at 37 kDa.

ab8069 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab8069 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 μg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (<a href='/ko/products/primary-antibodies/vimentin-antibody-v9-cytoskeleton-marker-ab8069'>ab8069</a>) at 1 µg/mL

Lane 1:

U-2 OS (Human bone osteosarcoma epithelial cell line) cell lysate at 20 µg

Lane 2:

Human tonsil cell lysate at 20 µg

Lane 2:

Western blot - Human VIM (Vimentin) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-vim-vimentin-knockout-hela-cell-line-ab255446'>ab255446</a>)

Lane 3:

Wild-type HeLa cell lysate at 20 µg

Lane 4:

VIM knockout HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 50 kDa,53 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human VIM (Vimentin) knockout HeLa cell lysate (AB263775)

Homozygous : Insertion of the selection cassette in exon2