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Description
Rat TNNI1 ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of standard gradient working solution: Add 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Troponin I Type 1, Slow Skeletal (TNNI1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the level of Troponin I Type 1, Slow Skeletal (TNNI1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Troponin I Type 1, Slow Skeletal ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Skeletal muscle slow-twitch troponin I (TNNI1) is a protein encoded by the TNNI1 gene. It is a tissue-specific isoform of troponin I, which is part of the troponin complex. TNNI functions to control the contraction and relaxation of skeletal muscle. Troponin I interacts with all major regulatory proteins in the sarcolemmal thin filaments of cardiac and skeletal muscle: troponin C, troponin T, myosin, and actin. When cell membrane Ca2+ levels are low, TNNI binds to the thin filaments, blocking the myosin binding sites on actin. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 31.25-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, and other biological fluids |
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4.5 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Inexpensive doesn’t mean cheap.
Color: Black/Orange
After 10 months, zero complaints. Well, I got rid of the horrible NATO strap 🙄. That’s a personal preference. Easy to swap out to whatever you like. Keeps accurate time meaning it’s the same time. I’m not micro analyzing it and the seconds gained/lost. It has a confident look and feel and makes you feel good when wearing it. Sure it’s not high priced but it’s not cheap. Very comfortable, light and not unincumbered. If this is what you’re looking at, you’d be satisfied. Great looking, durable and simple watch. Full confidence.
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Reviewed in the United States on October 25, 2025
★★★★★ 3
Budget Seiko Prospex this is not
Color: Black/Orange
Bought this as a cheap work watch, but I guess all the budget went into the solar technology.
No date adjust feature, gotta crank the crown over and over to adjust the date. (Also no screw-down crown, but that's expected at this price)
Fake diver bezel. Lol why? Cringe as heck. Should of just put a compass bezel on there instead tbh.
Cheap plastic crystal, scratches and mars just by looking at it.
Okay NATO strap. Falls apart quickly, but it's easily replaceable with a better quality one.
Decent lume on the hands, but none on the hour markers even though it looks like there is.
Looks cool.
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Reviewed in the United States on June 13, 2025
★★★★★ 5
Simplicity
Color: Black/Orange
This watch was received exactly as advertised. First off the watch is economical compared to other watches that advertise the same details. I will say that I have gone through numerous Luminox watches only to be disappointed. This watch fits the bill. It does have solar cells which mean it needs to be exposed to light to keep running, not a problem. Three minutes under my surefire light I am golden. The bezel doesn't rotate actually no big deal for me. I have always required that the bezel moves not anymore after having a bezel that rotates I use it next to never. Here's why, the more I rotate the bezel the more the numbers wear off. The strap on the watch with the catches for the strap are perfect. They actually hold the watch strap in place in lieu of like other big name brands that just flop and slide. As far as everyday use goes this watch is my go to. To be honest I have purchased three. This is the only watch I will wear moving forward, yeah the glass my scratch but even the big names like the Luminox I spoke of earlier scratches too.
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Well made
Size: 15.5" Neck 32"-33" Sleeve, Color: Tomato
Great color, nice quality, fits as expected
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Reviewed in the United States on May 4, 2023
★★★★★ 4
Great material. Sadly no pocket
The fit is not tailored. So if you are slim around the waist it will feel baggy. But enough material to tuck in or go around a large waist. The material is not stiff. Wash and dried on warm. This has been worn 2 times. If you take it out of dryer and hang up, the wrinkles come right out! The only drawback: No pocket. My wife didn't notice when she ordered, we threw package away otherwise would return, because it was a little pricey. But nice clean look!
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Reviewed in the United States on April 14, 2022
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