Anti-HspBP1 antibody (ab3858)

ICC/IF image of ab3858 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab3858, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in MCF7 cells.

Western blot using ab3858 on 20 ug per lane of MCF-7 Whole Cell Lysate. Secondary: Goat anti-rabbit IgG HRP conjugate ab6721 (1/2000). Exposure time: 2 mins.

Lane 1: ab3858 at 1/500.

Lane 2: ab3858 at 1/1000.

Paraffin embedded sections of human kidney tissue were incubated with ab3858 (1/75 dilution) at room temperature for 30 mins. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab3858 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org