**Determination of C-Peptide in Serum, Plasma and Related Liquid Samples of Mice**
(This kit is intended for in vitro research purposes only and is not suitable for clinical diagnosis.)
Disclaimer: Dear customers, thank you for choosing our products. This kit is manufactured using high-quality raw materials from renowned global suppliers and is produced using advanced ELISA technology. It is designed for the quantitative detection of natural and recombinant C-Peptide in mouse serum, plasma, tissue homogenates, or cell culture supernatants. Please read the instructions thoroughly before use and verify all reagent components. If you have any questions, feel free to contact Shanghai Jinma Biotechnology Co., Ltd. promptly.
**1. Mouse C-Peptide (C-Peptide) Quantitative Detection Kit (ELISA)**
**2. Required Experimental Equipment:**
- 37°C incubator
- Microplate reader with a 450 nm wavelength
- Precision pipettes and disposable tips
- Disposable test tubes
- Absorbent paper
**Procedure:**
1. **Preparation:** Remove the kit from the refrigerator and allow it to equilibrate at room temperature for 30 minutes.
2. **Dilution:** Dilute the 20× washing solution with distilled water to prepare the working solution.
3. **Sample and Standard Addition:** Use a microplate with designated wells for standards, samples, and blank controls. Add 50 μL of standard sample to each standard well, 10 μL of sample followed by 40 μL of diluent (total 5× dilution), and no sample to the blank control.
4. **Incubation:** Incubate the plate at 37°C for 30 minutes.
5. **Washing:** Discard liquid, pat dry, and wash each well with washing solution. Repeat this process four times.
6. **Enzyme Conjugate Addition:** Add 50 μL of enzyme conjugate to each well (except the blank control).
7. **Second Incubation:** Incubate again at 37°C for 30 minutes.
8. **Second Washing:** Wash the plate as described above.
9. **Color Development:** Add 50 μL of developer A, then 50 μL of developer B. Mix gently for 30 seconds and incubate at 37°C for 15 minutes.
10. **Stop Reaction:** Add 50 μL of stop solution to each well to terminate the reaction.
11. **Measurement:** Read the absorbance at 450 nm within 15 minutes of stopping the reaction.
12. **Data Analysis:** Plot the standard curve using standard concentrations and corresponding OD values. Calculate the sample concentration based on its OD value. Multiply by the dilution factor to obtain the actual concentration.
**Precautions:**
1. Avoid using samples containing sodium azide (NaN₃), as it may inhibit horseradish peroxidase (HRP).
2. Process samples immediately after collection. Store at -20°C if not tested right away; avoid repeated freeze-thaw cycles.
3. Centrifuge samples thoroughly to remove hemolysis or particulates.
4. Follow the manual strictly and rely on microplate reader readings for accurate results.
5. Seal unused enzyme conjugate in a desiccant bag immediately after opening.
6. Perform duplicate tests for standards, samples, and blanks to minimize errors.
7. Remember that the sample was diluted 5×, so multiply the result by 5 for the actual concentration.
8. The detection range is 62.5–2000 pg/mL. For values outside this range, dilute the sample with the provided diluent and retest.
9. If the color is too light, extend the substrate incubation time slightly.
10. Prevent cross-contamination by using separate tips and avoiding contact between reagents and microwells. Do not reuse sealing films.
11. Use the kit within the shelf life and do not mix reagents from different batches.
12. Protect Substrate B from light exposure.
**Mouse C-Peptide (C-Peptide) Quantitative Detection Kit**
This product introduces advanced, convenient, and efficient experimental technology from abroad. It offers fast, simple, accurate, and highly sensitive detection, with quick delivery and high-quality performance. Ready-to-use kits are available for immediate application in your research.
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