Lane 1 : Anti-HspBP1 antibody (ab3858) at 1/500 dilutionLane 2 : Anti-HspBP1 antibody (ab3858) at 1/1000 dilutionLane 1 : MCF-7 Whole Cell LysateLane 2 : MCF-7 Whole Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Image courtesy of Human Protein AtlasParaffin 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
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.