Immunofluorescence - Anti-HSP60 Rabbit pAb

Sample: Hela cells

The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.5% Triton X-100 for 10 minutes and then blocked with 5% BSA in 0.1% PBS-Tween for 0.5 hours.

Primary antibody: P900009 at 1:100 dilution

Secondary antibody: Goat anti-Rabbit (488) at 1:1,000 dilution (shown in green)

Nuclei were stained with DAPI (shown in blue).

All lanes: HSP60 Rabbit pAb at 1:1,000 dilution

Lane 1: Hep-G2 cell Lysates

Lane 2: Hela cell Lysates

Lane 3: MCF-7 cell Lysates

Lysates/proteins at 20 μg per lane.

Secondary

Goat Anti-Rabbit IgG (H+L), Peroxidase conjugated (LF102) at 1:5,000 dilution.

Observed band size: 63 kDa

Blocking/Dilution buffer: 1×PBST.

Immunohistochemical analysis of paraffin-embedded colon cancer tissue using Anti-HSP60 Rabbit pAb. Antigen repair using EDTA antigen repair solution and blocking with 5% BSA for 0.5 hour. Samples were incubated with primary antibody (1:150) overnight at 4℃. A undiluted HRP-labeled anti-rabbit IgG was used as the secondary antibody for 0.5 hour.

Immunohistochemical analysis of paraffin-embedded mouse skin tissue using Anti-HSP60 Rabbit pAb. Antigen repair using EDTA antigen repair solution and blocking with 5% BSA for 0.5 hour. Samples were incubated with primary antibody (1:150) overnight at 4℃. A undiluted HRP-labeled anti-rabbit IgG was used as the secondary antibody for 0.5 hour.

Immunohistochemical analysis of paraffin-embedded rat testis tissue using Anti-HSP60 Rabbit pAb. Antigen repair using EDTA antigen repair solution and blocking with 5% BSA for 0.5 hour. Samples were incubated with primary antibody (1:150) overnight at 4℃. A undiluted HRP-labeled anti-rabbit IgG was used as the secondary antibody for 0.5 hour.

Immunofluorescence (Formalin/PFA-fixed cells) - Anti-HSP60 antibody

Immunofluorescence analysis of HeLa cells labelling HSP60.

Cells were incubated overnight at room temperature with primary antibody (HSP60) at 2 µg/mL.

Secondary antibodies Goat anti-Rabbit (488) (shown in green) were used, incubated 1 h at room temperature.

DAPI was used as nuclear counterstain (shown in blue).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins.