Western blot - cardiac Troponin I antibody (ab47003)

All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1 µg/ml

Lane 1 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)
Lane 2 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 24 kDa
Observed band size : 26 kDa (why is the actual band size different from the predicted?)

Western blot - Cardiac Troponin I antibody (ab47003)

All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1 µg/ml

Lane 1 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)
Lane 2 : Heart (Mouse) Tissue Lysate
Lane 3 : Heart (Rat) Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 24 kDa
Observed band size : 26 kDa (why is the actual band size different from the predicted?)
Additional bands at : 160 kDa,80 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 3 minutes

Western blot - Cardiac Troponin I antibody (ab47003)

All lanes : Anti-Cardiac Troponin I antibody (ab47003) at 1/20000 dilution

Lane 1 : Pig Heart whole cell lysate
Lane 2 : Pig Heart whole cell lysate
Lane 3 : Pig Heart whole cell lysate
Lane 4 : Pig Heart whole cell lysate

Lysates/proteins at 4 µg per lane.

Secondary
An HRP-conjugated Goat anti-rabbit polyclonal at 1/20000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 24 kDa
Observed band size : 29 kDa (why is the actual band size different from the predicted?)


Blocking Step: 20% Serum for 3 hours at 24°C

This image is courtesy of an Abreview submitted by Dr. Mario Torrado

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cardiac Troponin I antibody (ab47003)

IHC image of Cardiac Troponin I staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab47003, 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.

Immunocytochemistry/ Immunofluorescence - Anti-Cardiac Troponin I antibody (ab47003)

ab47003 staining Cardiac Troponin I in Human cardiomyocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 20ºC. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for 1 hour at 20ºC. An undiluted FITC-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

This image is courtesy of an anonymous Abreview