重组Anti-Hsp90 beta抗体[E296] (ab32568)

Lanes 1-4: Merged signal (red and green). Green - ab32568 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab32568 Anti-Hsp90 beta antibody [E296] 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.

Immunohistochemical staining of paraffin embedded human stomach with purified ab32568 at a working dilution of 1 in 150. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

Immunofluorescence staining of HepG2 cells with purified ab32568 at a working dilution of 1 in 100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 555 goat anti rabbit (ab150082), used at a dilution of 1 in 400. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab32568 was used at a dilution of 1/200 followed by an Alexa Fluor^® 488 goat anti-mouse antibody at a dilution of 1/500.

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.

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Blocking buffer: 5% NFDM/TBST

Dilution buffer: 5% NFDM/TBST

Immunohistochemical analysis of paraffin-embedded human urinary bladder carcinoma using unpurified ab32568 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.