**Human GSTM1 ELISA Kit – For the quantitative in vitro determination of Human Glutathione S-Transferase mu 1 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and OTHER biological fluids. Intended for laboratory research use only. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE.**
This kit is designed to measure the levels of Human GSTM1 using an enzyme-linked immunosorbent assay (ELISA) method. The principle of the test involves a colorimetric reaction that is quantified at 450 nm using a spectrophotometer. A set of calibration standards is included to generate a standard curve, which allows the accurate determination of GSTM1 concentrations in unknown samples by comparing their optical density (OD) values.
**Intended Use:**
The Human GSTM1 ELISA Kit is intended for research purposes only and is not suitable for diagnostic or clinical applications. It is used to quantify GSTM1 levels in various biological samples such as serum, plasma, body fluids, and tissue homogenates.
**Sample Collection and Storage:**
- **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Remove serum and assay immediately or store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Use heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g (2–8°C). Store at -20°C if not used immediately.
- **Cell culture supernatants, tissue homogenates, and other biological fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Avoid repeated freezing and thawing.
*Note: Ensure proper centrifugation and avoid hemolysis or granulation in the samples.*
**Materials Required but Not Supplied:**
1. Incubator at 37°C
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**Reagents Provided:**
- Microtiter Strip Plate (12×8 or 12×4 strips)
- Standards (6 vials, 0.5 ml/vial)
- Sample Diluent (6.0 ml or 3.0 ml)
- HRP-Conjugate Reagent (10.0 ml or 5.0 ml)
- 20X Wash Solution (25 ml or 15 ml)
- Chromogen Solutions A & B (6.0 ml or 3.0 ml each)
- Stop Solution (6.0 ml or 3.0 ml)
- Closure Plate Membrane (2 units)
- User Manual (1 copy)
- Sealed Bags (1 unit)
**Standard Concentrations:**
1200, 600, 300, 150, 75, 37.5 pg/mL. If sample values exceed the highest standard, dilute with Sample Diluent. Ensure all reagents are from the same kit lot.
**Important Instructions:**
1. Do not substitute reagents between different kits.
2. Allow all reagents to reach room temperature (20–25°C) before use. Do not use water baths for thawing.
3. Do not use reagents past their expiration date.
4. Use only deionized or distilled water for dilutions.
5. Keep microtiter plates in sealed bags until ready for use. Unused strips should be stored at 2–8°C with desiccant.
6. Use fresh pipette tips for each transfer to prevent cross-contamination.
7. Handle all disposable materials as potentially hazardous. Wear gloves during the procedure.
8. Waste must be inactivated for at least 30 minutes before disposal.
9. Substrate solution is sensitive to contamination; discard if it appears bluish.
10. Chromogen B contains 20% acetone; keep away from heat or flame.
11. Allow all reagents to reach room temperature before starting the assay.
**Reagent Preparation:**
- **Wash Solution (1X):** Mix 1 volume of 20X Wash Solution with 19 volumes of distilled/deionized water. Store at 2–8°C for up to one month.
**Assay Procedure:**
1. Prepare all reagents before starting. Add 50 µL of standard or sample in duplicate to the microtiter plate.
2. Add 100 µL of HRP-Conjugate Reagent to all wells except the blank. Cover and incubate for 60 minutes at 37°C.
3. Wash the plate four times manually or automatically.
4. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
5. Add 50 µL of Stop Solution to each well. The color should turn yellow. If green or uneven, gently mix.
6. Measure OD at 450 nm. Generate a standard curve by plotting average OD vs. concentration.
7. Determine sample concentrations by comparing their OD values to the standard curve.
**Performance Characteristics:**
- Intra-assay CV <15%, Inter-assay CV <15%
- Detection range: 37.5 pg/mL – 1200 pg/mL
- Sensitivity: <1.0 pg/mL
- Cross-reactivity: No significant interference observed
**Storage Conditions:**
- Store at 2–8°C for frequent use.
- Store at -20°C for long-term storage (up to 6 months).
**Notes:**
- Always perform a standard curve for each run.
- Variations in technique, timing, or environmental conditions may affect results.
- Follow all safety guidelines when handling biological samples and reagents.
**For Research Use Only.**
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