ab93837 (1/200) staining EWSR1 in assynchronous HeLa Cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.See Abreview
All lanes : Anti-EWSR1 antibody (ab93837) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 3 : A498 (Human Kidney Carcinoma) Whole Cell LysateLane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell LysateLane 5 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 7 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ICC/IF image of ab93837 stained HepG2 cells. 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 (ab93837, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) 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 antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.
IHC image of EWSR1 staining in Human normal testis 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 ab93837, 1µg/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.