**Quantitative Determination of C-Peptide in Mouse Serum, Plasma, and Related Biological Samples**
(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 ELISA kit is manufactured using high-quality raw materials from renowned global suppliers and follows advanced production 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.
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### **Product Description**
1. **Mouse C-Peptide (C-Peptide) Quantitative ELISA Kit**
2. **Required Equipment for Experimentation**
- 37°C incubator
- Microplate reader with 450 nm wavelength
- Precision pipettes and disposable tips
- Disposable test tubes
- Absorbent paper
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### **Experimental Procedure**
1. **Preparation**
Remove the kit from the refrigerator and allow it to equilibrate at room temperature for 30 minutes.
2. **Dilution of Washing Solution**
Dilute the 20× concentrated washing solution with distilled water to prepare the working solution.
3. **Sample and Standard Addition**
- Use a microplate with pre-labeled wells: standard wells, sample wells, and blank control.
- Add 50 μL of standard solution to each standard well.
- Add 10 μL of the sample to be tested, followed by 40 μL of sample diluent (total 5× dilution).
- The blank control well receives no sample.
4. **Incubation**
Incubate the plate at 37°C for 30 minutes.
5. **Washing**
Discard the liquid, pat dry on absorbent paper, then fill each well with washing solution. Let stand for 1 minute, discard, and repeat this process 4 times. Alternatively, use an automated washer as instructed.
6. **Add Enzyme-Linked Standard Working Solution**
Add 50 μL of enzyme-linked standard working solution to each well, except the blank control.
7. **Second Incubation**
Incubate again at 37°C for 30 minutes.
8. **Second Washing**
Repeat the washing procedure as above.
9. **Color Development**
Add 50 μL of Developer A, followed by 50 μL of Developer B. Mix gently for 30 seconds and incubate at 37°C for 15 minutes. Avoid exposure to light during this step.
10. **Stop Reaction**
Add 50 μL of stop solution to each well. The color will change from blue to yellow.
11. **Measurement**
Measure the absorbance (OD value) at 450 nm within 15 minutes after stopping the reaction. Use the blank well for zeroing.
12. **Data Analysis**
Plot a standard curve using the OD values and corresponding concentrations. Calculate the sample concentration based on the regression equation. You may also use software for analysis. Multiply the result by the dilution factor to obtain the actual sample concentration.
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### **Precautions and Tips**
1. Avoid using samples containing sodium azide (NaN₃), as it can inhibit horseradish peroxidase (HRP).
2. Process samples as soon as possible after collection. If not used immediately, store at -20°C and avoid repeated freeze-thaw cycles.
3. Ensure samples are fully centrifuged and free from hemolysis or particulates.
4. Follow the manual strictly. Results must be confirmed using a microplate reader.
5. Store unused enzyme reagents in sealed bags with desiccant.
6. It is recommended to run standards and samples in duplicate to reduce error.
7. Remember that the sample was diluted 5×; multiply the final result by 5 to get the true concentration.
8. The detection range is 62.5–2000 pg/mL. For results outside this range, adjust by diluting with the provided diluent and re-testing.
9. If the color development is too weak, extend the substrate incubation time slightly.
10. Prevent cross-contamination by using separate pipette tips for each addition. Avoid touching the microwells with common reagents. Do not reuse the sealing film.
11. Use the kit within its shelf life and do not mix reagents from different batches.
12. Developer B is light-sensitive—keep it away from direct light.
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### **Product Features**
This Mouse C-Peptide Quantitative ELISA Kit introduces advanced, convenient, and efficient experimental technology from abroad. It offers fast, simple, accurate, and highly sensitive detection, with short delivery times and excellent quality. It is available for immediate shipment.
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