Human hypoglycosylated IgA1 (UGIGA1) elisa detection kit instructions - Database & Sql Blog Articles

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Kaixin micro test

Test probe P100-M3

Brand AVX TPSE226M035R0125 Low impedance tantalum capacitor AVX 22

Human hypoglycosylated IgA1 (UGIGA1)

ELISA Test

Kit Instructions

This kit is for research use only and not intended for human or animal diagnostic purposes.

Experimental Principle

The ELISA kit utilizes a double-antibody sandwich assay to quantify the level of human hypoglycosylated IgA1 (UGIGA1) in biological samples. The microplate is pre-coated with a specific antibody against UGIGA1, which captures the target antigen from the sample. After incubation, an HRP-conjugated secondary antibody binds to the captured antigen, forming an immune complex. Following washing steps, TMB substrate is added, producing a blue color that turns yellow upon addition of stop solution. The intensity of the color is directly proportional to the concentration of UGIGA1 in the sample. The 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.

Human Hypoglycosylated IgA1 (UGIGA1)

ELISA Test

Kit Composition

1. 130x Washing Solution – 20ml × 1 bottle

2. Stop Solution – 6ml × 1 bottle

3. Enzyme Standard Reagent – 6ml × 1 bottle

4. Standard (16μg/ml) – 0.5ml × 1 bottle

5. Enzyme-labeled Coating Plate – 12 wells × 8

6. Sample Diluent – 6ml × 1 bottle

7. TMB Substrate A – 6ml × 1 bottle

8. TMB Substrate B – 6ml × 1 bottle

9. Standard Dilutions – 1.5ml × 1 bottle

10. Instruction Manual – 1 copy

11. Sealing Film – 2 sheets

12. Sealed Bag – 1

Sample Requirements

1. Samples should be processed as soon as possible after collection. If testing cannot be done immediately, store at -20°C. Avoid repeated freeze-thaw cycles.

2. Samples containing NaN3 are not suitable for this assay, as sodium azide may inhibit horseradish peroxidase (HRP) activity.

ELISA Procedure

1. Standard Dilution: Prepare serial dilutions of the original standard according to the provided table.

2. Loading: Add 50 μl of standard, 40 μl of sample diluent, and 10 μl of sample into each well. Ensure accurate pipetting and gentle mixing.

3. Incubation: Seal the plate and incubate at 37°C for 30 minutes.

4. Washing: Use 30x diluted washing buffer (prepared with distilled water) and wash the plate 5 times.

5. Enzyme Addition: Add 50 μl of enzyme-labeled reagent to each well except blank controls.

6. Incubation: Repeat the 37°C incubation for 30 minutes.

7. Color Development: Add 50 μl of TMB A and 50 μl of TMB B, incubate at 37°C for 10 minutes.

8. Stop Reaction: Add 50 μl of stop solution to each well to terminate the reaction.

9. Reading: Measure OD values at 450 nm within 15 minutes of stopping the reaction.

Calculation

Plot a standard curve using known concentrations and corresponding OD values. Calculate the unknown sample concentration based on the curve. Multiply the result by the dilution factor to obtain the actual concentration.

Precautions

1. Allow the kit to reach room temperature before use. Store unopened enzyme reagents in a sealed bag.

2. If washing solution crystallizes, warm it gently in a water bath before use.

3. Use a dedicated pipette and ensure accuracy. Limit loading time to 5 minutes if possible.

4. Always prepare a standard curve and run duplicates. For high-concentration samples, perform a preliminary dilution.

5. Use a new sealing film for each experiment to prevent cross-contamination.

6. Protect the substrate from light during storage and handling.

7. Follow all instructions carefully to ensure accurate results.

8. Treat all waste materials as biohazardous.

9. Do not mix components from different batches.

Storage Conditions & Expiration

1. Store the kit at 2–8°C.

2. Shelf life: 6 months from the date of manufacture.