人GPI (AMF) knockout HEK-293T cell裂解物

3 Images

• WB

Lab

Western blot - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : GPI knockout HEK293T cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab167394 was shown to specifically react with AMF in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266834 (knockout cell lysate ab257458) was used. Wild-type and AMF knockout samples were subjected to SDS-PAGE. ab167394 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-AMF antibody [EPR11663(B)] (<a href='/products/primary-antibodies/amf-antibody-epr11663b-ab167394'>ab167394</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

GPI knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human GPI (AMF) knockout HEK-293T cell line (<a href='/products/cell-lines/human-gpi-amf-knockout-hek-293t-cell-line-ab266834'>ab266834</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 63 kDa

Observed band size: 63 kDa

false

• WB

Lab

Western blot - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)

Lane 1 : wild-type HEK-293T cell lysate 20 ug
Lane 2 : GPI knockout HEK-293T cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab66340 observed at 63 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab66340 was shown to react with AMF in wild-type HEK-293T cells in Western blot with loss of signal observed in GPI knockout cell line ab266834 (GPI knockout cell lysate ab257458). Wild-type HEK-293T and GPI knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab66340 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 ^°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-AMF antibody [1B7D7] (<a href='/products/primary-antibodies/amf-antibody-1b7d7-ab66340'>ab66340</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

GPI knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human GPI (AMF) knockout HEK-293T cell line (<a href='/products/cell-lines/human-gpi-amf-knockout-hek-293t-cell-line-ab266834'>ab266834</a>)

Predicted band size: 63 kDa

Observed band size: 63 kDa

false

• Sanger seq

Unknown

Sanger Sequencing - Human GPI (AMF) knockout HEK-293T cell lysate (AB257458)

Homozygous : 76 bp deletion in exon2