Method for making homocysteine assay kit

The production methods of homocysteine assay kits mainly include two types: enzyme cycling assay kits and ELISA assay kits. The following are the production steps and key points:

Enzyme cycling assay kit production method

First reagent preparation:

Raw material preparation: According to the ratio requirements, add 100mmol/L buffer system, preservative, and 0.01-0.06g/L serine into the mixing container.

pH adjustment: Adjust the pH value of the mixing container to 9.0 ± 0.2.

. Addition of reducing agent: Add 10-100mmol/L of reducing agent (such as TCEP, GPC, EGTA, etc.) to the mixing container, stir and let it stand, then filter.

Second raw material stirring: Stir the mixing container at the set stirring time and speed.

. Place the mixed container at 2-8 degrees Celsius for 24 hours after mixing, and filter it using stainless steel filters and filter membranes. Add the second raw material (such as NADH 0.1-0.5g/L and LDH 10-50ku/L) to the filtered mixing container and stir to obtain the first reagent.

Second reagent preparation:

Raw material preparation: According to the ratio requirements, add the obtained buffer system 50mmol/L, stabilizer 1-100g/L, and preservative into another mixing container.

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pH adjustment: Adjust the pH value of the mixing container to 7.6 ± 0.2.

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Fourth raw material mixing: Add the fourth raw material (such as cystathionine β - synthase 1-10KU/L, cystathionine β - lyase 1-10KU/L, activator 0.001-0.01g/L, etc.) to the mixing container, and mix to obtain the second reagent.

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ELISA kit preparation method

Preparation of pre coated plate:

Antibody coating: Use purified homocysteine to capture the antibody and coat the microporous plate to make solid-phase antibodies.

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Preparation of enzyme labeled antibody working solution:

Enzyme labeling: HRP (horseradish peroxidase) is labeled onto the detection antibody to prepare an enzyme labeled antibody working solution.

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Standard preparation:

Standard preparation: Prepare standard solutions containing different concentrations of homocysteine.

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Preparation of other reagents:

Preparation of colorimetric solution: Prepare TMB (tetramethylbenzidine) colorimetric solution.

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Preparation of termination solution: Prepare a termination solution for the termination reaction (such as 1N sulfuric acid).

. Preparation of washing solution: Prepare a washing solution for washing plates.