Goat IgM ELISA Kit
2025-10-06 23:08:59
**Goat IgM ELISA Kit – For the quantitative in vitro determination of Goat Immunoglobulin M concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only, not for diagnostic or therapeutic purposes. Please read this entire package insert before using the product.**
**INTENDED USE AND TEST PRINCIPLE**
This Goat IgM ELISA Kit is designed for laboratory research applications and is not suitable for use in diagnostic or clinical procedures. The kit utilizes an enzyme-linked immunosorbent assay (ELISA) to quantitatively measure IgM levels in various biological samples. The color change from blue to yellow, caused by the Stop Solution, allows for the measurement of optical density (OD), which correlates with IgM concentration. A set of calibration standards is included to generate a standard curve, enabling accurate quantification of IgM 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, then centrifuge at 2000×g for 20 minutes. 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 of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid multiple freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids**: Remove particulates by centrifugation. Assay immediately or aliquot and store at -20°C. Ensure proper centrifugation and avoid hemolysis or contamination.
**MATERIALS REQUIRED BUT NOT SUPPLIED**
1. 37°C incubator
2. Microplate reader capable of measuring absorbance at 450 nm
3. Pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**REAGENTS PROVIDED**
All reagents should be stored at 2–8°C. Check expiration date on label.
| Reagent Name | 96 Determinations | 48 Determinations |
|---------------------------|-------------------|-------------------|
| MicroELISA Strip Plate | 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 |
**NOTES**
1. Standard concentrations: 48, 24, 12, 6, 3, 1.5 μg/mL.
2. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
3. Do not thaw samples or reagents using water baths.
4. Do not use reagents beyond the expiration date.
5. Only use deionized or distilled water for reagent dilution.
6. Keep microtiter plate in the sealed bag until use. Unused strips must be stored with desiccant at 2–8°C.
7. Use fresh pipette tips for each transfer to prevent cross-contamination.
8. Disposable knives should not be used in the assay due to potential risk of infectious agents.
9. All samples should be disposed of following biosafety protocols.
10. Liquid waste should be treated with 1% sodium hypochlorite for at least 30 minutes before disposal.
11. Substrate solution is sensitive; if it appears bluish, discard.
12. Chromogen B contains 20% acetone—keep away from heat or flame.
13. Allow all reagents to reach room temperature (20–25°C) before use.
**REAGENT PREPARATION AND STORAGE**
- **Wash Solution (1X)**: Mix 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to 1 month.
**ASSAY PROCEDURE**
1. Prepare all reagents before starting. It is recommended to 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 blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the microtiter plate 4 times (manual or automated).
- *Manual*: Aspirate, fill with 1X Wash Solution, aspirate again. Repeat 4 times. Invert and blot dry.
- *Automated*: Aspirate and wash four times with 350 μL/well. Invert and blot dry.
5. 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.
6. Add 50 μL of Stop Solution to each well. Read OD at 450 nm within 15 minutes.
**CALCULATION**
1. Plot average OD (450 nm) against standard concentrations on a graph.
2. Subtract blank OD values before interpretation.
3. Construct the standard curve using graph paper or statistical software.
4. At the point of intersection, draw a vertical line to the X-axis to determine IgM concentration.
5. Results may vary due to operator technique, pipetting accuracy, washing efficiency, or kit age. Each user should establish their own standard curve.
6. Intra-assay CV: <15%, Inter-assay range: 1.5–48 μg/mL.
7. Sensitivity: <1.0 μg/mL.
8. No significant cross-reactivity observed.
9. Storage: 2–8°C (frequent use); 6 months at -20°C.
**WARNING**
Do not use this kit for diagnostic or clinical testing. Always follow safety guidelines when handling biological materials.
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