L-Malic Acid Kit Recommendation For Microplate Format:
Either K-LMALAF or K-LMALQR are recommended for use in a 96-well microplate format and the main advantages/disadvantages are described below:
K-LMALAF and K-LMALQR kits are designed for use in the auto-analyser format and therefore can be used without any modification to assay volumes directly in a 96-well microplate format.
These kits assays have less reagent additions than K-LMAL-58A/116A.
K-LMALQR is designed for use in the auto-analyser format and is identical in performance to K-LMALAF except that the reagents are supplied ready prepared so no reagents preparation is required.
K-LMALAF or K-LMALQR kits do not have MegaCalc applications available to enable easy results calculation which therefore must be achieved as outlined above in either of points 2 or 3.
If preferred, K-LMAL-58A/116A may also be easily converted for use in a 96-well microplate format. Basically, the assay volumes for the cuvette format must be reduced approximately 10-fold for use in a 96-well microplate. However, some assay optimisation may be required (e.g. increased enzyme concentration etc.) and unlike the cuvette which has a set path-length of 1 cm, the path-length in the microplate is dependent upon the volume of liquid in the well.
K-LMAL-58A/116A assay volumes used in the manual assay format should be reduced by 10-fold for use in a 96-well microplate format (some assay optimisation may be required, e.g. increased enzyme concentration etc.).
The calculation of results is achieved as outlined below in either of points 1, 2 or 3.
The main advantage here is that if this kit is used with a microplate reader that has a path-length conversion capability, or if results are converted as outlined above in point 3, then this enables easy calculation of results using the K-LMAL-58A/116A MegaCalc application (available on the Megazyme website where the product is located).
To enable the calculation of the amount of analyte in the samples from tests performed in the microplate format one of the following must be done:
1. The easiest method is to use a microplate reader that has a path-length conversion capability (i.e. the microplate reader can detect the path-length of each well and convert the individual readings to a 1 cm path-length). This will allow values to be calculated using the MegaCalc calculation software which can be found where the product is located on the Megazyme website.
2. Perform a standard curve of the analyte on each microplate that contains test samples and calculate the result of the test samples from the calibration curve (concentration of analyte versus absorbance).
3. Perform a standard curve of the analyte in both the cuvette format (i.e. with a 1 cm path-length) and the 96-well microplate format and use these results to obtain a mean conversion factor between the cuvette values and the microplate values.