Anti-Cardiac Troponin I antibody [EP1106Y] (ab52862) at 1/1000000 dilution + fetal heart cell lysate at 10 µg

Secondary
Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 28 kDa
Observed band size: 28 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cardiac muscle tissue sections labeling Cardiac Troponin I with Purified ab52862 at 1:300 dilution (1.19 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

ab52862 (purified) at 1:30 dilution (2µg) immunoprecipitating Cardiac Troponin I in Human fetal heart lysate.
Lane 1 (input): Human fetal heart lysate 10µg
Lane 2 (+): ab52862 & Human fetal heart lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52862 in Human fetal heart lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.

IHC image of Cardiac Troponin I staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab52862, 1/200 dilution, 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

Flow cytometry analysis of A-673 (Human muscle Ewing's Sarcoma ) cells labeling Cardiac Troponin I (red) with ab52862 at a 1/30 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

ab52862 at 1/250 dilution staining cardiac Troponin I in human cardiac muscle by Immunohistochemistry, Paraffin embedded tissue.