Human IL18 knockout HeLa cell lysate

3 Images

• WB

Lab

Western blot - Human IL18 knockout HeLa cell lysate (AB256952)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : IL18 knockout HeLa cell lysate (20 μg)

Lane 3 : A431 cell lysate (20 μg)

Lane 4 : Jurkat cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab207324 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.

ab207324 Anti-IL-18 antibody [EPR19954] was shown to specifically react with IL-18 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265274 (knockout cell lysate ab256952) was used. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. ab207324 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 200 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-IL-18 antibody [EPR19954] (<a href='/en-us/products/primary-antibodies/il-18-antibody-epr19954-ab207324'>ab207324</a>) at 1/200 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

IL18 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human IL18 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-il18-knockout-hela-cell-line-ab265274'>ab265274</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

• WB

Lab

Western blot - Human IL18 knockout HeLa cell lysate (AB256952)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : IL18 knockout HeLa cell lysate (20 μg)

Lane 3 : A431 cell lysate (20 μg)

Lane 4 : Jurkat cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab243091 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.

ab243091 Anti-IL-18 antibody [EPR19954-188] was shown to specifically react with IL-18 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265274 (knockout cell lysate ab256952) was used. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. ab243091 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) 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 (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-IL-18 antibody [EPR19954-188] (<a href='/en-us/products/primary-antibodies/il-18-antibody-epr19954-188-ab243091'>ab243091</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

IL18 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human IL18 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-il18-knockout-hela-cell-line-ab265274'>ab265274</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human IL18 knockout HeLa cell lysate (AB256952)

Homozygous : 1 bp deletion in exon4