TNNI3 ELISA 试剂盒

This assay has high sensitivity and excellent specificity for detection of High Sensitive Troponin I Type 3, Cardiac (TNNI3).
No significant cross-reactivity or interference between High Sensitive Troponin I Type 3, Cardiac (TNNI3) and analogues was observed.

• Pre-coated, ready to use 96-well strip plate
• Plate sealer for 96 wells
• Standard Diluent
• Assay Diluent A
• Assay Diluent B
• Stop Solution
• Standard
• Detection Reagent A
• Detection Reagent B
• TMB Substrate
• Wash Buffer (30 x concentrate)
• Instruction manual

• Microplate reader with 450 nm filter.
• Precision single or multi-channel pipettes and disposable tips.
• Eppendorf Tubes for diluting samples.
• Deionized or distilled water.
• Absorbent paper for blotting the microtiter plate.
• Container for Wash Solution

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit is not used up all at once, remove only the strips and reagents for the current experiment and leave the remaining strips and reagents in the desired condition.
2. Standard - Reconstitute the Standard with 1.0mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 800 pg/mL.First dilute the stock solution to 200 pg/mL and the diluted standard serves as the highest standard (200 pg/mL). Then prepare 7 tubes containing 0.5mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 200pg/mL, 100pg/mL, 50pg/mL, 25pg/mL,12.5pg/mL, 6.25pg/mL, 3.12pg/mL, and the last tube with Standard Diluent is the blank as 0pg/mL
3. Detection Reagent A and Detection Reagent B - Spin or centrifuge the stock of Detection Reagent A and B briefly before use. Dilute to working concentration (1:100) with Assay Diluent A or B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution Concentrate (30x) with 580 mL of deionized or distilled water to make 600 mL of Wash Solution (1x).
5. TMB Substrate - Aspirate the required amount of solution with sterile tip and do not return the residual solution back into the vial.

Note:

1. Serial dilution directly in the wells is not recommended.
2. Prepare standard within 15 minutes before assay. Do not dissolve the reagents directly at 37 °C.
3. Detection Reagent A and B are sticky solutions, so pipette them slowly to reduce volume errors.
4. Reconstitute Standard or working solutions of Detection Reagent A and B carefully according to instructions, avoiding foaming and mixing gently until crystals are completely dissolved. To minimize inaccuracy caused by pipetting, use small volumes and ensure pipettes are calibrated. It is recommended to aspirate more than 10 µL for one-time pipetting.
5. The reconstituted Standard, Detection Reagent A and B can only be used once.
6. When crystals have formed in the Wash Solution concentrate (30x), warm it to room temperature and mix gently until the crystals are completely dissolved.
7. Contaminated water or preparation containers affect the detection result.

1. Prepare all reagents, samples and standards,
   
2. Add 50μL standard or sample to each well.
   And then add 50μL prepared Detection Reagent A immediately.
   Shake and mix. Incubate 1 hour at 37 °C,
   
3. Aspirate and wash 3 times,
   
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
   
5. Aspirate and wash 5 times,
   
6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
   
7. Add 50μL Stop Solution. Read at 450 nm immediately.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Troponin I Type 3, Cardiac (TNNI3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Troponin I Type 3, Cardiac (TNNI3) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.