Western blot - Hsp90 antibody [AC88] (ab13492)

All lanes : Anti-Hsp90 antibody [AC88] (ab13492) at 1/1000 dilution

Lane 1 : Hsp90 native human protein
Lane 2 : Hsp90 beta reombinant human protein
Lane 3 : Hsp90 alpha reombinant human protein
Lane 4 : Cell lysates prepared from heat shocked Hela cells
Lane 5 : Cell lysates prepared from heat shocked 3T3 cells
Lane 6 : Cell lysates prepared from heat shocked PC-12 cells
Lane 7 : Cell lysates prepared from heat shocked CHO-K1 cells
Lane 8 : Cell lysates prepared from heat shocked Rat-2 cells


Predicted band size : 84.7 (alpha) , 83.2 (beta) kDa

Immunocytochemistry - Hsp90 antibody [AC88] (ab13492)

ab13492 staining Hsp90 in cultured human AGS cells by Immunocytochemistry. Cells were fixed with formaldehyde, permeabilized with 0.025% Triton-X in TBS and blocking with 5% serum was performed for 1 hour at 23⁰C. Samples were incubated with primary antibody (1/100) for 1 hour at 23°C. An undiluted HRP-conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

This image is a courtesy of Anonymous Abreview

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Hsp90 antibody [AC88] (ab13492)

ab13492 staining Hsp90 in human stomach tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocking with 5% serum for 1 hour at 23°C was performed. The primary antibody was diluted 1/100 and incubated with sample for 1 hour at 23°C. A HRP conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

This image is a courtesy of Anonymous Abreview

Immunoprecipitation - Hsp90 antibody [AC88] (ab13492)

ab13492 Immunoprecipitate OF hSP 90 in human AGS whole cell lysate. 200µg of cell lysate was incubated with primary antibody (1/250 in RIPA buffer) and matrix (Protein A/G) for 16 hours at 4ºC.
For western blotting Ab6728 (1/1000) was used.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Hsp90 antibody [AC88] (ab13492)

ICC/IF image of ab13492 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13492, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.