Troponin I Type 3, Cardiac (TNNI3) BioAssay™ ECL Kit (Human)
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Troponin I Type 3, Cardiac (TNNI3) BioAssay™ ECL Kit (Human) is a quantitative, in vitro, Chemiluminescence-based sandwich immunoassay (ECL) suitable for use in Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids.
The microtiter plate provided in this kit has been pre-coated with an antibody specific to Troponin I Type 3, Cardiac. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Troponin I Type 3, Cardiac. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Troponin I Type 3, Cardiac (TNNI3) level in the sample or standard.
The minimum detectable dose of this kit is typically less than 10.5pg/ml.
This sandwich assay has high sensitivity and excellent specificity for detection of Troponin I Type 3, Cardiac (TNNI3). No significant cross-reactivity or interference between Troponin I Type 3, Cardiac (TNNI3) and analogues was observed.
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Microtiter Plate, 1x96 wells, Pre-coated, ready to use, well strip plate
Plate sealer for 96 wells
Standard, 2x1 vial
Standard Diluent, 1x20ml
Detection Reagent A, 1x120ul
Detection Reagent B, 1x120ul
Assay Diluent A, 1x12ml
Assay Diluent B, 1x12ml
Substrate A, 1x10ml
Substrate B, 1x2ml
Wash Buffer, 30X, 1x20ml
Materials Required but not Supplied:
1. Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0secs; read time 1.0 sec/well.
2. Precision single or multi-channel pipettes and pipette tips with disposable tips.
3. Eppendorf Tubes for diluting samples.
4. Deionized or distilled water.
5. Absorbent paper for blotting the microtiter plate.
6. Container for Wash Solution
1. Prepare all reagents, samples and standards.
2. Add 100ul standard or sample to each well. Incubate 2 hours at 37°C.
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C.
4. Aspirate and wash 3 times.
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C.
6. Aspirate and wash 5 times.
7. Add 100ul Substrate Solution. Incubate 10 minutes at 37°C.
8. Read RLU value immediately.
Use a serum separator tube and allow samples to clot for two hours at RT or overnight at 4°C before centrifugation for 20 minutes at approximately 1000×g. Assay freshly prepared serum immediately or store samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2-8°C within 30 minutes of collection.
Remove plasma and assay immediately or store samples in aliquots at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles.
Other Biological Fluids:
Centrifuge samples for 20 minutes at 1000xg. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C for later use.
Avoid repeated freeze/thaw cycles.
Samples to be used within 5 days may be stored at 4°C, otherwise samples must be stored at -20°C ( 1 month) or -80°C ( 2 months) to avoid loss of bioactivity and contamination.
Sample hemolysis will influence the result, so hemolytic specimens cannot be detected.
When performing the assay, bring samples to room temperature.
Storage and Stability:
1. 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.
2. 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.
3. Expiration date is 6 months from shipment date.
4. It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.