ab111447 stained MCF-7 cells. The cells were 100% methanol fixed (5 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 ab111447 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
Anti-Cytokeratin 10 antibody (ab111447) at 1 µg/ml + Skin (Human) Tissue Lysate - adult normal tissue (ab30166) at 10 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
IHC image of Cytokeratin 10 staining in human skin melanoma 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 ab111447, 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.
IHC-P image of Cytokeratin 10 staining using human skin using ab111447 (1:6000). The sections were subjected to heat mediated antigen retrieval using citric acid pH 6. The sections were blocked using 1% BSA for 10 mins at 21°C. ab111447 was incubated for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to biotin (1:250).See Abreview
IHC-P image of Cytokeratin 10 staining using rat E18 skin using ab111447 (1:5000). The sections were subjected to heat mediated antigen retrieval using citric acid pH 6. The sections were blocked using 1% BSA for 10 mins at 21°C. ab111447 was incubated for 2 hours at 21°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to biotin (1:250).See Abreview