ab13492 staining Hsp90 (red) and another antibody to C2GnT-M (Golgi enzyme, greeen) in Panc-1 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with PFA and blocked with 1% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/50 1% Donkey serum in PBST) for 1 hour at 22°C. An undiluted DyLight® 594-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody.

This image was generated using the ascites version of the product. 

See Abreview

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

Lane 2 : HSP90 cell lysate
Lane 3 : HSP90ß cell lysate
Lane 4 : HSP90a cell lysate
Lane 5 : HeLa (heat shocked) cell lysate
Lane 6 : NIH/3T3 (Heat shocked) cell lysate
Lanes 7-8 : PC-12 (Heat shocked) cell lysate

This image was generated using the ascites version of the product.

Anti-Hsp90 antibody [AC88] (ab13492) at 1/1000 dilution + Human fibroblast whole cell lysate at 40 µg

Secondary
HRP-conjugated goat anti-mouse polyclonal IgG at 1/2000 dilution

Developed using the ECL technique.

Exposure time: 20 seconds

Blocked with 5% Milk for 2 hours at 22°C

This image was generated using the ascites version of the product.

See Abreview

ab13492 staining Hsp90 in Human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a TRIS-EDTA Buffer. Samples were incubated with primary antibody (1/500) for 30 minutes at 20°C. A HRP-conjugated Goat anti-rabbit/mouse IgG polyclonal was used as the secondary antibody.

This image was generated using the ascites version of the product.

See Abreview

Overlay histogram showing HeLa cells stained with ab13492 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13492, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

ab13492 staining Hsp90 in Human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (10 ug/ml).

This image was generated using the ascites version of the product.

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

This image was generated using the ascites version of the product.

ICC/IF image of ab13492 stained HepG2 cells (ab7900). 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 +4°C. 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.

This image was generated using the ascites version of the product.