Jan 16, 2026
Standard Operating Procedure (SOP): Mouse Troponin T Type 2 (cardiac) ELISA Kit (Colorimetric)
- Elizabeth Rocker1
- 1The Jackson Laboratory
- CARDIOVERSE
Protocol Citation: Elizabeth Rocker 2026. Standard Operating Procedure (SOP): Mouse Troponin T Type 2 (cardiac) ELISA Kit (Colorimetric). protocols.io https://dx.doi.org/10.17504/protocols.io.e6nvwnd1dvmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: January 14, 2026
Last Modified: January 16, 2026
Protocol Integer ID: 238638
Keywords: troponin isoform, troponin complex, tropomyosin, biological marker for cardiomyocyte, indicator of myocardial cell, myocardial cell, biological marker, cardiac, cardiomyocyte, calcium sensitivity, elisa kit, contractile apparatus in muscle
Abstract
Troponin T is one of three troponin isoforms found in the tropomyosin-troponin complex. This complex is responsible for the calcium sensitivity of the contractile apparatus in muscle. Cardiac Troponin T is used as a biological marker for cardiomyocytes and its level in serum is frequently used as an indicator of myocardial cell injury.
Troubleshooting
Administrative
Version:
1.0
Effective
Date: 01/05/2026
Approved
by: Beth Rocker, Senior Research Project Manager
Prepared
by: Rileah Slayton, Research Assistant II
Sample Preparation
Dilute the experimental samples (serum, plasma, lysates or other biofluids) and perform a
serial dilution of the standard (purified form of the analyte under
consideration) in Reagent Diluent. If available, use the literature to
determine the optimal concentration and linear working range. Samples should be
prepared in duplicate, or preferably in triplicate.
Procedure
Use an ELISA compatible PVC clear microtiter plate (96-well) and determine the number of wells needed for the assay.
Dispense 100 µL of Capture Antibody Solution into the wells. Apply sealing tape to the top of the plate to prevent evaporation. Incubate the plate overnight at 4 °C. After incubation remove the tape and aspirate each well.
Dispense 400 µL of Wash Buffer into each well. Then invert the plate between washes and flick out the solution. Wash the plate 2 more times and pat the inverted plate on a paper towel to dry.
Block plates by dispensing 300 µL of Reagent Diluent into each well and incubate the plate for 60 minutes at room temperature. After incubation, remove the tape. Invert the plate and flick out the solution.
Repeat step 3.
Add 100 µL of diluted samples and standards to the appropriate wells and cover the plate.
Incubate the plate for 2 hours at room temperature. After incubation, uncover the plate and aspirate each well.
Repeat step 3.
Dispense 100 µL of Detection Antibody Solution to each well. Incubate for 2 hours at room temperature and cover the plate. After incubation, uncover the plate. Invert the plate and flick out the solution.
Repeat step 3.
Dispense 100 µL of SA Solution into each well. Incubate for 20 minutes at room temperature and cover the plate. After incubation, uncover the plate. Invert the plate and flick out the solution.
Repeat step 3.
Dispense 100 µL of the Substrate Solution into each well. Cover the plate and incubate for to 30 minutes at room temperature. Avoid placing the plate in direct sunlight.
TIP: If no color develops within 30 minutes, incubation time can be extended until an appropriate color change/signal is acquired. TMB substrate reacts with HRP to generate a blue colored product which changes to yellow upon the addition of Stop Solution, whereas the PNPP substrate reacts with AP to generate a yellow-colored product.
After color development, remove the cover and dispense 50 µL of Stop Solution into each well to stop the enzymatic reaction.
Immediately measure the absorbance of each well using a spectrophotometer/plate reader with the appropriate absorbance setting.
TIP: The presence of air bubbles in wells may impact the results and it is advisable to spin the plate in a centrifuge.