Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue labelling USP10 with ab70895 at 1/1000 (0.2µg/ml). Detection: DAB.
All lanes : Anti-USP10 antibody (ab70895) at 0.04 µg/mlLane 1 : Whole cell lysate from HeLa cells at 50 µgLane 2 : Whole cell lysate from HeLa cells at 15 µgLane 3 : Whole cell lysate from HeLa cells at 5 µgLane 4 : Whole cell lysate from 293T cells at 50 µg
Detection of Human USP10 by Immunoprecipitation in Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab70895 at 3 µg/mg lysate for IP, and at 1 µg/ml for subsequent Western blot detection.
All lanes : Anti-USP10 antibody (ab70895) at 0.1 µg/mlLane 1 : TCMK-1 whole cell lysateLane 2 : 4T1 whole cell lysateLane 3 : CT26.WT whole cell lysateLysates/proteins at 50 µg per lane.developed using the ECL technique
ICC/IF image of ab70895 stained HeLa cells. The cells were 4% formaldehyde 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 (ab70895, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab70895 staining in human normal tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70895, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.