Human glycophosphatidylinositol (GPI) ELISA kit instruction manual - Huaqiang Electronic Network

EL-C1600N100013-B

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Kaixin micro test

Test probe P100-M3

Kit

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Human Glycophosphatidylinositol (GPI) ELISA Kit

User's Guide

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Kit Name

Human Glycophosphatidylinositol (GPI) ELISA Kit

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Kit Use]

Quantitative detection of GPI in serum, plasma, and related fluid samples.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Detection Principle

This kit uses a two-antibody sandwich ELISA method. Standards and samples are added to pre-coated plates with enzyme-labeled antibodies. After incubation, unbound components are washed away. Substrates A and B are then added, producing a blue color that turns yellow under acidic conditions. The intensity of the color is proportional to the GPI concentration, measured at 450 nm. The sample’s GPI content is calculated based on the standard curve.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Kit Composition

Notes: Standard solutions should be diluted to 200, 100, 50, 25, 12.5, 6.25 ng/ml.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Reagents and Equipment Required but Not Provided

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1. Incubator at 37°C

2. Microplate reader

3. Precision pipettes and disposable tips

4. Distilled water

5. Disposable test tubes

6. Absorbent paper

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Procedure

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1. Preparation: Bring the kit to room temperature for 30 minutes after removing from the refrigerator.

2. Dilute the 20× washing solution to the working concentration using distilled water.

3. Add standards and samples: Place the enzyme-labeled plate on the frame. Set up standard, sample, and blank control wells. Add 50 μL of standard to each standard well, 10 μL of sample and 40 μL of diluent to each sample well, and no addition to the blank control.

4. Incubate at 37°C for 30 minutes.

5. Wash the plate 4 times with washing solution, ensuring complete removal of liquid and drying on absorbent paper.

6. Add 50 μL of enzyme standard working solution to each well except the blank control.

7. Incubate again for 30 minutes at 37°C.

8. Repeat step 5 for washing.

9. Add 50 μL of substrate A, then 50 μL of substrate B. Incubate in the dark at 37°C for 15 minutes.

10. Add 50 μL of stop solution to each well to terminate the reaction. The color will change from blue to yellow.

11. Measure OD values at 450 nm within 15 minutes after stopping the reaction.

12. Calculate the standard curve and determine the sample concentration using linear regression. Multiply by the dilution factor to obtain the actual concentration.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Sample Requirements

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1. Avoid sodium azide (NaN₃) in samples, as it inhibits horseradish peroxidase (HRP).

2. Extract samples immediately after collection, and store at -20°C if not tested right away. Avoid repeated freeze-thaw cycles.

3. Centrifuge samples thoroughly to remove hemolysis and particles.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Precautions

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1. Follow the manual strictly and rely on microplate reader readings for results.

2. Store unused enzyme-labeled reagents in sealed bags with desiccants.

3. Perform duplicate tests for standards, samples, and blanks to reduce errors.

4. Remember to multiply the result by 5 if the sample was diluted fivefold.

5. The detection range is 0.1–200 ng/mL. If outside this range, dilute the sample appropriately and recalculate.

6. If the color is too light, extend the substrate incubation time slightly.

7. Prevent cross-contamination by using new pipettes and tips for each addition. Do not reuse any reagents or seal films.

8. Use the kit within its shelf life and do not mix reagents from different batches.

9. Protect Substrate B from light exposure.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Summary of Procedures

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Prepare reagents, samples, and standards → Add samples and standards → Incubate at 37°C for 30 min → Wash 4 times → Add enzyme reagent → Incubate 30 min → Wash 4 times → Add substrates A and B → Incubate 15 min → Add stop solution → Read OD values within 15 min → Calculate results.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Detection Range

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0.1–200 ng/mL

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Specification

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96T/box, 48T/box

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Storage

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Store at 2–8°C, protected from light and moisture.

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Human Glycophosphatidylinositol (GPI) ELISA Kit

Shelf Life

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6 months