Lane 1: Wild-type HEK-293T cell lysate (20µg)

Lane 2: PRKCSH knockout HEK-293T cell lysate (20µg)
Lane 3: HeLa cell lysate (20µg)
Lane 4: Daudi cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab129098 observed at 80 kDa. Red - loading control ab8245 observed at 37 kDa.
ab129098 Anti-Glucosidase 2 subunit beta antibody [EPR8047] was shown to specifically react with PRKCSH in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266770 (knockout cell lysate ab257608) was used. Wild-type and PRKCSH knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab129098 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 10000 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.

Lane 1: Wild-type HEK-293T cell lysate (20µg)

Lane 2: PRKCSH knockout HEK-293T cell lysate (20µg)
Lane 3: HeLa cell lysate (20µg)
Lane 4: Daudi cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab134071 observed at 80 kDa. Red - loading control ab8245 observed at 37 kDa.
ab134071 Anti-Glucosidase 2 subunit beta antibody [EPR8046] was shown to specifically react with PRKCSH in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266770 (knockout cell lysate ab257608) was used. Wild-type and PRKCSH knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134071 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.

Allele-1: 14 bp deletion in exon 1

 

Allele-2: 13 bp deletion in exon 1

 

Allele-3: Insertion of the selection cassette in exon 1

 

Allele-4: Insertion of the selection cassette in exon 1