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TNNI3 ELISA Kit

TNNI3 Reactivity: Dog Colorimetric Sandwich ELISA
Catalog No. ABIN956369
  • Target See all TNNI3 ELISA Kits
    TNNI3 (Cardiac Troponin I (TNNI3))
    Reactivity
    • 6
    • 6
    • 6
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    Dog
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Analytical Method
    Quantitative
    Characteristics
    The Dog Cardiac Troponin-I, High Sensitive ELISA uses two affinity purified antibodies. A rabbit anti-cTnI polyclonal antibody is used for solid phase immobilization (on the microtiter wells). A goat anti-cTnI peptide-specific polyclonal antibody is conjugated to horseradish peroxidase (HRP) and used for detection. Samples (serum or plasma) and calibrators (200 µL) are pipetted into the microtiter wells and incubated for 2 hours on a plate shaker. After washing the wells, 100 µL of diluent and 100 µL of HRP-conjugated anti-cTnI are pipetted into each of the microtiter wells. The plate is incubated for one hour on a plate shaker. During this step, cTnI becomes sandwiched between the solid phase and HRP-conjugated antibodies. The wells are then washed to remove unbound HRP-conjugated antibodies. TMB, an HRP substrate (100 µL), is then added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped by the addition of 1N HCl (100 µL), changing the color to yellow, and absorbance at 450 nm is measured. The concentration of cTnI is proportional to the absorbance at 450 nm and is derived from a calibration curve.
    Components
    Anti-cTnI-coated microtiter plate (96 wells 12 x 8-well strips)
    cTnI Stock: Lyophilized dog cTnI
    cTnI Diluent (25 mL)
    cTnI HRP Conjugate (11 mL)
    20X Wash Solution (50 mL)
    TMB Reagent (11 mL): HRP substrate solution
    Stop Solution (11 mL): 1N HCl.
    Material not included
    Distilled or de-ionized water
    Pipettes & tips: P-10, P-200 & P-1000 or equivalent
    Plate shaker
    Plate washer
    Plate reader capable of reading OD at 450 nm
    Vortex mixer
    Absorbent paper
    Graph paper or appropriate PC graphing software
    Polypropylene microcentrifuge tubes (1.5 mL)
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  • Plate
    Pre-coated
    Assay Procedure
    1. Secure the desired number of coated wells in the holder.
      2. Dispense 200 µL of calibrators and samples into appropriate wells.
      3. Incubate on an orbital shaker (150 rpm) at 25°C for 2 hours.
      4. Using a plate washer, wash the microtiter wells 5 times with 1X wash solution (400 µL per well).The entire wash procedure should be performed as quickly as possible.
      5. Strike the wells sharply onto adsorbent paper or paper towels to remove all residual droplets.
      6. Add 100 µL of cTnI diluent to each well.
      7. Add 100 µL of HRP conjugate to each well.
      8. Incubate on an orbital shaker (150 rpm) at 25°C for 1 hour.
      9. Wash the plate as described in steps 4 and
      5.
      10. Dispense 100 µL of TMB Reagent into each well.
      11. Incubate on an orbital shaker (150 rpm) at 25°C for 20 minutes.
      12. Stop the reaction by adding 100 µL of Stop Solution to each well.
      13. Gently mix until all the blue color changes to yellow.
      14. Read absorbance at 450 nm with a plate reader within 5 minutes.
      15. If absorbance values exceed the high calibrator, the samples should be further diluted with cTnI diluent and re-determined.
    Calculation of Results
    1. Calculate the mean absorbance value (A450) for the calibrators and samples.
      2. Construct a calibration curve by plotting the A450 values obtained for each reference calibrator against its concentration in ng/mL on graph paper, with absorbance on the vertical (y) axis and concentration on the horizontal (x) axis.
      3. Using the A450 values for each sample, determine the corresponding concentration of cTnI (ng/mL) from the calibration curve.
      4. If available, graphing software should be used to analyze the data. Depending on the range of the calibration curve used for analysis, good fits may be obtained with linear regression analysis or by fitting the data to a polynomial second order equation. We recommend the latter option.
    Restrictions
    For Research Use only
  • Storage
    -20 °C
    Storage Comment
    When the kit is received, store the lyophilized calibrator at or below -20°C. Store the remainder of the kit in the refrigerator at 4°C. Keep the microtiter plate in a sealed bag with desiccant. The kit expiration date is indicated on the box label.
    Expiry Date
    The expiry date is stated on the label.
  • Target See all TNNI3 ELISA Kits
    TNNI3 (Cardiac Troponin I (TNNI3))
    Alternative Name
    Cardiac Troponin-I, High Sensitive (TNNI3 Products)
    Synonyms
    CMD1FF ELISA Kit, CMD2A ELISA Kit, CMH7 ELISA Kit, RCM1 ELISA Kit, TNNC1 ELISA Kit, cTnI ELISA Kit, CMD1Z ELISA Kit, CMH13 ELISA Kit, TN-C ELISA Kit, TNC ELISA Kit, TNNC ELISA Kit, Tn1 ELISA Kit, TnIc ELISA Kit, ctnIc ELISA Kit, XTnIc ELISA Kit, c-troponin ELISA Kit, cTNT ELISA Kit, cmh7 ELISA Kit, ctni ELISA Kit, tnnc1 ELISA Kit, TNNI3 ELISA Kit, TnI ELISA Kit, cTNI ELISA Kit, AI874626 ELISA Kit, TnC ELISA Kit, cTnC ELISA Kit, tncc ELISA Kit, Tncc ELISA Kit, troponin I3, cardiac type ELISA Kit, troponin C1, slow skeletal and cardiac type ELISA Kit, troponin I, cardiac 3 ELISA Kit, troponin I type 3 (cardiac) ELISA Kit, troponin I3, cardiac type S homeolog ELISA Kit, troponin I3, cardiac type L homeolog ELISA Kit, cardiac troponin I ELISA Kit, troponin C, cardiac/slow skeletal ELISA Kit, TNNI3 ELISA Kit, TNNC1 ELISA Kit, Tnni3 ELISA Kit, tnni3.S ELISA Kit, tnni3.L ELISA Kit, tnni3 ELISA Kit, LOC100462680 ELISA Kit, Tnnc1 ELISA Kit
    Background
    Troponin is the contractile regulating protein complex of striated muscle. It consists of three subunits: troponin I, T, and C. Troponin-I exists in three isoforms, one in fast-twitch skeletal muscle, one in slow-twitch skeletal muscle, and one in cardiac muscle. After muscle injury by trauma or ischemia, troponin is released into the bloodstream and its levels correlate well with tissue injury. The sequence of cardiac troponin-I (cTnI) is significantly different from the skeletal muscle isoforms, thus allowing generation of specific anti-cTnI antibodies and development of cTnI specific immunoassays. Because cTnI is uniquely expressed in the heart it provides a specific biomarker of cardiac damage.
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