ab1385 staining human tonsil by IHC-P.
ab1385 staining Cytokeratin 15 in human normal skin tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking in 10% serum for 30 minutes at 21°C. The primary antibody was diluted, 1/25 in PBT (PBS + 0.5% Tween-20 + 0.5% BSA) and incubated with sample for 30 minutes at 21°C. A HRP conjugated goat polyclonal to mouse IgG1 was used undiluted as secondary.See Abreview
ICC/IF image of ab1385 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 (ab1385, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.
Overlay histogram showing A431 cells stained with ab1385 (red line). The cells were fixed with 80% methanol (5 min) and and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1385, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed.
ab1385 staining Cytokeratin 15 in formalin-fixed, paraffin-embedded Human skin tissue by Immunohistochemistry. Staining was detected using DAB.