**Human MFG-E8 ELISA Kit – For the quantitative in vitro determination of Human Milk Fat Globule EGF Factor 8 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other biological fluids. FOR LABORATORY RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE.** This ELISA kit is designed for research purposes only and should not be used in diagnostic or therapeutic procedures. The assay relies on a colorimetric reaction where the Stop Solution changes the color from blue to yellow. The optical density (OD) is measured at 450 nm using a spectrophotometer. To accurately quantify MFG-E8 levels in your samples, the kit includes a set of calibration standards. These standards are run alongside the samples to generate a standard curve, which is essential for determining the concentration of MFG-E8 in unknown samples by comparing their OD values to the curve. --- **Sample Collection and Storage:** - **Serum:** Use a serum separator tube. Allow samples to clot 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 aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000 ×g (2–8°C). Avoid freezing and thawing multiple times. - **Cell culture supernatants, tissue homogenates, and other body fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present in the samples. --- **Materials Required (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 (Stored at 2–8°C):** | Component | 96 Determinations | 48 Determinations | |----------|-------------------|-------------------| | MicroELISA Stripplate | 12×8 strips | 12×4 strips | | Standard (6 vials) | 0.5 ml/vial | 0.5 ml/vial | | Sample Diluent | 6.0 ml | 3.0 ml | | HRP-Conjugate Reagent | 10.0 ml | 5.0 ml | | 20X Wash Solution | 25 ml | 15 ml | | Chromogen Solution A | 6.0 ml | 3.0 ml | | Chromogen Solution B | 6.0 ml | 3.0 ml | | Stop Solution | 6.0 ml | 3.0 ml | | Closure Plate Membrane | 2 | 2 | | User Manual | 1 | 1 | | Sealed Bags | 1 | 1 | *Standard concentrations: 240, 120, 60, 30, 15, 7.5 pg/mL.* *If sample values exceed the highest standard, dilute with Sample Diluent and retest.* --- **Precautions:** 1. Do not mix reagents from different kit lots. All components are calibrated for optimal performance. 2. Allow all reagents to reach room temperature (20–25°C) before use. Avoid using water baths for thawing. 3. Do not use reagents past their expiration date. 4. Only use deionized or distilled water for dilutions. 5. Store unused strips in their sealed pouch with desiccant at 2–8°C. 6. Use fresh pipette tips for each transfer to prevent cross-contamination. 7. Handle all blood-derived samples with care, as they may pose infection risks. 8. Wear gloves during the procedure. 9. Dispose of waste properly after inactivation (allow to stand for 30 minutes). 10. Substrate solutions must be handled carefully; discard if discolored. 11. Substrate B contains 20% acetone—keep away from heat or flame. 12. Allow all reagents to warm to room temperature before starting the assay. --- **Reagent Preparation:** - **Wash Solution (1X):** Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month. --- **Assay Procedure:** 1. Prepare all reagents before beginning. Run standards and samples in duplicate. 2. Add 50 µL of standard or sample to appropriate wells. Blank well receives no addition. 3. Add 100 µL of HRP-Conjugate Reagent to all wells except the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times. Manual washing involves aspirating and refilling with Wash Solution. Automated washing requires careful control of fill volume. 5. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protecting from light. 6. Add 50 µL of Stop Solution to each well. The color should turn yellow. If green or uneven, gently tap the plate. 7. Measure OD at 450 nm. Generate a standard curve by plotting average OD values against standard concentrations. --- **Data Interpretation:** - Calculate the mean OD for each standard and sample. Subtract blank OD from all values. - Plot the standard curve and determine sample concentrations by interpolation. - Intra-assay and inter-assay CVs are <15%. - Detection range: 7.5 pg/mL to 240 pg/mL. - Sensitivity: <1.0 pg/mL. - Cross-reactivity: No significant cross-reactivity observed with human MFG-E8. **Storage:** Store at 2–8°C for frequent use; for long-term storage, keep at -20°C for up to six months. **Note:** Always follow the manufacturer's instructions and maintain good laboratory practices.

Toilet Plunger & Brush

Toilet Plunger,Push Toilet Clog Cleaning Tools,Toilet Clog Remover,Toilet Brush

ZHEJIANG HAOCHUAN RUBBER &PLASTIC CO.,LTD , https://www.zjhaochuan.com