Instructions for using homocysteine assay kit

The usage of homocysteine (HCY) assay kit varies depending on the specific assay method and kit type. The following are two common methods for using homocysteine assay kits:

Enzyme cycle assay kit

Reagent and sample preparation

Reagent: Ensure that Reagent 1 and Reagent 2 are within their expiration dates and stored at 2-8 ℃ according to the instructions. The opened reagent can remain stable for 30 days in the instrument compartment (2-8 ℃).

Sample: Collect fasting serum or plasma as a sample. The sample should be transported and stored under low temperature conditions. After collection, it should be refrigerated and quickly centrifuged. The sample can be stored for 48 hours at 2-8 ℃. If the test is delayed, the serum needs to be refrigerated.

sample processing

If the HCY content in the sample exceeds 50 μ mol/L, it should be diluted with 8.5% saline and measured, and the result should be multiplied by the dilution factor.

According to the requirements of different instruments, the dosage of reagents and samples can be adjusted proportionally. The usual dosage for reagent 1 is 185 μ L, for reagent 2 it is 50 μ L, and for the sample it is 10 μ L.

Instrument settings

Using a fully automatic biochemical analyzer, set the measurement temperature to 37 ℃, wavelength to 340nm, colorimetric cup diameter to 1.0cm, measurement mode to rate method, and measurement time to 120 seconds.

Operation steps

Add reagent 1 and reagent 2 to the instrument according to the instructions.

Add the sample to the instrument and begin the measurement.

The instrument will automatically complete the reaction and detection process and provide measurement results.

Result analysis

Based on the measurement results, combined with the standard curve or reference value range, determine the concentration of HCY in the sample.

If the measurement result exceeds the reference value range, it should be retested for confirmation.

elisa kit

Reagent and sample preparation

Reagents: Ensure that all reagents are within their expiration date and stored according to the instructions. Standard samples, biotin labeled antibody working solution, horseradish peroxidase labeled avidin working solution, etc. need to be prepared before use.

Sample: Collect serum, plasma, cell culture supernatant, etc. as samples, which should be stored under low temperature conditions to avoid repeated freezing and thawing.

sample processing

If the concentration of HCY in the sample is too high, dilute it with sample diluent to make the sample meet the detection range of the kit.

Sample addition and reaction

Set blank holes, standard holes, and test sample holes according to the instructions.

Add standard or test sample to each well separately, and then add biotinylated antibody working solution and horseradish peroxidase labeled avidin working solution.

Incubate the reaction plate in a 37 ℃ constant temperature incubator for a certain period of time.

Board washing and color rendering

Wash the board according to the instructions and remove unbound reagents.

Add color reagent to each well and incubate in the dark for a certain period of time.

Add termination solution to stop the reaction.

Result determination

Measure the optical density (OD value) of each well using an enzyme-linked immunosorbent assay (ELISA) reader at a wavelength of 450nm.

Draw a standard curve based on the OD value and concentration of the standard sample.

Find the corresponding concentration on the standard curve based on the OD value of the sample to be tested.

Result analysis

Based on the measurement results, determine the concentration of HCY in the sample.

If the measurement result exceeds the reference range, it should be retested for confirmation and combined with other clinical information for comprehensive analysis.

Please note that different brands and models of reagent kits may have differences in details, so it is important to carefully read the instructions and follow the instructions before use. At the same time, safety precautions should be taken during the experiment to avoid reagents splashing onto sensitive areas such as the skin or eyes.