The K-LACGAR assay is still compatible with 96-well microplate format by simply reducing the assay volumes recommended for the manual cuvette format by 10-fold. 

Note: You should be aware that, unlike the cuvette which has a set pathlength of 1 cm, the pathlength in the microplate is dependent upon the volume of liquid in the well. This means that the manual calculation given in the data booklet or the MegaCalc software can not be used when using the microplate format.


1) The easiest method is to use a microplate reader that has a path-length conversion capability (i.e. the microplater 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. Subsequent assays in the microplate format can then be converted from the calculated conversion factor.

We have not tested this assay in microplate format so the above guidelines are a suggestion and must be validated internally. We recommend that the kit control is run to determine if the method is working correctly.