Anti-Hsp90 alpha antibody [68/Hsp90]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody [68/Hsp90] (AB282108)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Hsp90 alpha with ab282108 at 1/50 (19.98 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in HeLa cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 alpha antibody [68/Hsp90] (AB282108)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Hsp90 alpha with ab282108 at 1/50 (19.98 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in 293T cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• WB

Lab

Western blot - Anti-Hsp90 alpha antibody [68/Hsp90] (AB282108)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 7 seconds

All lanes:

Western blot - Anti-Hsp90 alpha antibody [68/Hsp90] (ab282108) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 85 kDa

Observed band size: 83 kDa

false

• WB

Lab

Western blot - Anti-Hsp90 alpha antibody [68/Hsp90] (AB282108)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1-3 : Merged signal (red and green). Green - ab282108 observed at 83KDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab282108 Anti-Hsp90 alpha antibody was shown to react with Hsp90 alpha in HEK293T cells in Western blot. Loss of signal was observed when HSP90AA1 knockout sample was used. Wild-type and
HSP90AA1 knockout samples were subjected to SDS-PAGE.

ab282108 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) preadsorbed (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Hsp90 alpha antibody [68/Hsp90] (ab282108) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HSP90AA1 KO HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 85 kDa

Observed band size: 83 kDa

false