ab10245 staining Cardiac Troponin C in Human sternal bone marrow derived MSCs induced to cardio by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 1 hour at 37°C. Samples were incubated with primary antibody (1/250 in PBS + 1% Goat serum) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.See Abreview
Ab10245 staining human heart (left ventricle). Staining is localized to the cytoplasm.Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffers citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.