Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3: Saos-2 cell lysate 20 ug
Lane 4: HL-60 cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab58950 observed at 85 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab58950 was shown to react with Hsp90 in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90AB1 knockout cell line ab266117 (HSP90AB1 knockout cell lysate ab257190). Wild-type and HSP90AB1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab58950 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4^°C at 0.5 ug/ml 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 hour at room temperature before imaging.

Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3: Saos-2 cell lysate 20 ug
Lane 4: HL-60 cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab53497 observed at 85 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab53497 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90AB1 knockout cell line ab266117 (HSP90AB1 knockout cell lysate ab257190). Wild-type and HSP90AB1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab53497 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4^°C at a 1 in 5000 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 hour at room temperature before imaging.

Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3: Saos-2 cell lysate 20 ug
Lane 4: HL-60 cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab32568 observed at 85 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab32568 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90AB1 knockout cell line ab266117 (HSP90AB1 knockout cell lysate ab257190). Wild-type and HSP90AB1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab32568 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C at a 1 in 200000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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.

Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: HSP90AB1 knockout HEK-293T cell lysate 20 ug
Lane 3: Saos-2 cell lysate 20 ug
Lane 4: HL-60 cell lysate 20 ug
Lanes 1 - 4:Merged signal (red and green). Green - ab203085 observed at 85 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab203085 was shown to react with Hsp90 beta in wild-type HEK-293T cells in western blot with loss of signal observed in HSP90AB1 knockout cell line ab266117 (HSP90AB1 knockout cell lysate ab257190). Wild-type and HSP90AB1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab203085 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4^°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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.

Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:HSP90AB1 knockout HEK293T cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:SH-SY5Y cell lysate (20 ug)

ab203085 was shown to specifically react with Hsp90 beta in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. ab203085 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.

Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:HSP90AB1 knockout HEK293T cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:SH-SY5Y cell lysate (20 ug)

ab32568 was shown to specifically react with Hsp90 beta in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266117 (knockout cell lysate ab257190) was used. Wild-type and Hsp90 beta knockout samples were subjected to SDS-PAGE. ab32568 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.