**Human Interleukin-35 (IL-35) ELISA Kit – Instructions for Use**
This kit is intended for research use only and is designed to quantitatively determine the concentration of human interleukin-35 (IL-35) in various biological samples, including serum, plasma, urine, cell culture supernatants, and tissue homogenates.
**Principle of Operation**
The IL-35 ELISA kit utilizes a sandwich immunoassay technique. A microtiter plate pre-coated with a specific anti-IL-35 antibody is used as the solid phase. After incubation with the sample, IL-35 binds to the immobilized antibody. An HRP-conjugated secondary antibody then recognizes the captured IL-35, forming an antibody-antigen-enzyme complex. Following washing steps, TMB substrate is added, which reacts with HRP to produce a blue color that turns yellow upon acid termination. The intensity of the color is directly proportional to the IL-35 concentration in the sample. Absorbance is measured at 450 nm using a microplate reader, and the concentration is determined by comparing the sample OD value to a standard curve.
**Kit Components**
- Microtiter Plate: 1 × 48 or 1 × 96 wells
- Standard: 0.5 mL × 1 vial (90 ng/L)
- Standard Diluent: 1.5 mL × 1 vial
- Enzyme Conjugate: 3 mL × 1 vial
- Sample Diluent: 3 mL × 1 vial
- TMB Substrate A & B: 3 mL × 1 vial each
- Stop Solution: 3 mL × 1 vial
- Wash Buffer (20×): 20 mL × 20 times or 20 mL × 30 times
- Sealing Film: 2 pieces (for 48-well) or 2 pieces (for 96-well)
- Storage: 2–8°C
**Sample Preparation**
- **Serum/Plasma**: Centrifuge at 2000–3000 rpm for 20 minutes after clotting or anticoagulant addition.
- **Urine**: Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant**: Centrifuge after collection; for intracellular components, lyse cells via freeze-thaw cycles before centrifuging.
- **Tissue Homogenate**: Weigh the tissue, homogenize in PBS, centrifuge, and collect the supernatant.
- **Storage**: Samples should be processed promptly. If not tested immediately, store at -20°C and avoid repeated freeze-thaw cycles.
**Procedure Summary**
1. Prepare serial dilutions of the standard.
2. Add samples and standards to the plate.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate five times with diluted wash buffer.
5. Add enzyme conjugate and incubate again.
6. Add TMB substrate and incubate for 15 minutes.
7. Stop the reaction with stop solution.
8. Measure absorbance at 450 nm.
9. Calculate concentrations using a standard curve.
**Notes and Precautions**
- Allow the kit to reach room temperature before use.
- Avoid cross-contamination by using separate tips for each step.
- Ensure accurate pipetting and consistent timing.
- Do not mix reagents from different batches.
- Store all waste materials as biohazardous.
- Keep the substrate away from light.
- Always perform a standard curve and consider sample dilution if OD values exceed the standard range.
**Calculation**
Plot the standard curve using standard concentrations and corresponding OD values. Determine the sample concentration from the curve and adjust for any dilution factor.
**Performance**
- Sensitivity: 3 ng/L to 70 ng/L
- Correlation coefficient (R²): ≥ 0.990
- Intra- and inter-assay variation: < 9% and < 11%, respectively
**Storage and Shelf Life**
- Store the kit at 2–8°C.
- Shelf life: 6 months from the date of manufacture.
**Important**
Follow the instructions carefully. Results must be based on microplate reader readings. For detailed guidance, refer to the accompanying English manual.
Bagasse Pulp Food Plate
Bagasse Pulp Container A disposable printing plate made from bagasse pulp, a by-product of sugarcane processing. Bagasse pulp is the fibrous material left after extracting the juice from the cane stems. Instead of being discarded as waste, bagasse is collected and used to make a variety of products, including dinner plates.
Bagasse pulp plate are an environmentally friendly alternative to single-use plates made from traditional plastic or styrofoam. They are biodegradable and compostable, which means they can be broken down naturally and returned to the environment without harm. This makes them a more sustainable option for food service establishments and individuals seeking to reduce their environmental impact.
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