TNNI3 Kit ELISA

N° du produit ABIN956369

Détail du produit TNNI3 Kit ELISA

Antigène: TNNI3 (Cardiac Troponin I (TNNI3))

Reactivité: Chien

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Analytical Method: Quantitative

Attributs du produit: 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.

Ingrédients: 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.

Matériel non inclus: 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)

Information d'application

Plaque: Pre-coated

Procédure de l'essai:

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.

Calcul des résultats:

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

Stockage

Stock: -20 °C

Stockage commentaire: 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.

Date de péremption: The expiry date is stated on the label.

Détail du antigène

Antigène: TNNI3 (Cardiac Troponin I (TNNI3))

Autre désignation: Cardiac Troponin-I, High Sensitive (TNNI3 Produits)

Synonymes: CMD1FF Kit ELISA, CMD2A Kit ELISA, CMH7 Kit ELISA, RCM1 Kit ELISA, TNNC1 Kit ELISA, cTnI Kit ELISA, CMD1Z Kit ELISA, CMH13 Kit ELISA, TN-C Kit ELISA, TNC Kit ELISA, TNNC Kit ELISA, Tn1 Kit ELISA, TnIc Kit ELISA, ctnIc Kit ELISA, XTnIc Kit ELISA, c-troponin Kit ELISA, cTNT Kit ELISA, cmh7 Kit ELISA, ctni Kit ELISA, tnnc1 Kit ELISA, TNNI3 Kit ELISA, TnI Kit ELISA, cTNI Kit ELISA, AI874626 Kit ELISA, TnC Kit ELISA, cTnC Kit ELISA, tncc Kit ELISA, Tncc Kit ELISA, troponin I3, cardiac type Kit ELISA, troponin C1, slow skeletal and cardiac type Kit ELISA, troponin I, cardiac 3 Kit ELISA, troponin I type 3 (cardiac) Kit ELISA, troponin I3, cardiac type S homeolog Kit ELISA, troponin I3, cardiac type L homeolog Kit ELISA, cardiac troponin I Kit ELISA, troponin C, cardiac/slow skeletal Kit ELISA, TNNI3 Kit ELISA, TNNC1 Kit ELISA, Tnni3 Kit ELISA, tnni3.S Kit ELISA, tnni3.L Kit ELISA, tnni3 Kit ELISA, LOC100462680 Kit ELISA, Tnnc1 Kit ELISA

Sujet: 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.