Human Cardiac Troponin I, cTnI ELISA Kit

Catalog No.: BEK1253

Size: 96T

Range: 312 pg/ml - 20,000 pg/ml

Sensitivity 10 pg/ml

Storage and Expiration: Store at 4℃ for 6 months, or at -20℃ for one year. Store HRP conjugated antibody at 4℃ for one year.

Application: For quantitative detection of human cTnI in serum, cell culture supernatants or tissue homogenate.

Introduction

Troponin I is one of 3 subunits that form the troponin complex of the thin filaments of striated muscle, the others are troponin T and troponin C. Human troponin I is presented in cardiac muscle tissue by a single isoform with molecular weight 23876 Da and it consists of 209 amino acid residues. It binds actin and inhibits actomyosin ATPase activity in the absence of calcium. In 2000, Labarrere et al. found that persistent elevation of cardiac troponin I levels after heart transplantation were associated with high risk for developing coronary artery disease and graft failure after cardiac transplantation. The cardiac troponin I may be a useful tool in identifying patients with heart failure who are at increased risk for progressive ventricular dysfunction and death.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-cTnI monoclonal antibody was pre-coated onto 96-well plates. The standards and test samples were added to the wells and any cTnI present is bound by the immobilized antibody. After washing away any unbound substances, the HRP conjugated anti-cTnI monoclonal antibody was added to wells as detection antibodies. Following a wash to remove any unbound antibody-enzyme reagent, the TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding stop solution. The density of yellow is proportional to the cTnI amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of cTnI can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human cTnI antibody
2. Lyophilized human cTnI standards: 2 tubes (20 ng / tube)                  
3. Sample / Standard diluent buffer: 30 ml
4. HRP conjugated anti-human cTnI antibody (Concentrated): 130 μl. Dilution: 1:100 (Store at 4℃, do NOT store it at -20℃)
5. Antibody diluent buffer: 12 ml
6. TMB substrate: 10 ml
7. Stop solution: 10 ml
8. Wash buffer: 30 ml (25x). Dilution: 1:25

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. Microplate reader (wavelength: 450nm)
2. Precise pipette and disposable pipette tips
3. Deionized or distilled water
4. Automated plate washer
5. Beakers, flasks or cylinders, etc.
6. Statistical calculator
7. Plate cover
8. Absorbent filter papers or absorbent material
9. ELISA shaker