National Institute for Occupational Safety and Health

“ Using a pipettor with aerosol resistant tips, in an orderly fashion, add 80 μL aliquots of the standards, samples, and controls to the microtiter plate wells in duplicate. NOTE: The ELISA assay is a time dependent matrix. Add reagents in a consistent, uninterrupted manner to avoid variability and a drift effect over the plate. d. In the same order the standards and samples were added to the plate, add 2 drops of enzyme conjugate from the enzyme conjugate bottle. NOTE: If more than 3 rows of wells are to be employed, use a multiwell pipettor. ”

“ However, the higher the methanol concentration of the samples, the more desirable it is to use the same % of methanol standards as in the samples. b. Using the 50,000 ng/L solution, prepare the working standards 100 ng/L, 500 ng/L and 25000 ng/L, in the matrix solvent. 9. Run controls, 1 for every three rows of the assay to determine possible drift. 10. Zero the reader against air or deionized water before reading the assay. 11. Determine the concentration of alachlor using the data reduction capabilities of the microtiter plate reader, or appropriate software. ”

“ Run the diluted samples in the assay. TM Follow the general instructions of the EnviroGard Alachlor plate kit [4] . Adjust as needed to accommodate sample needs. a. Determine the numberof microtiter plate wells needed to run in duplicate the kit negative control and 3 kit standards, the matrix negative control and 3 matrix standards, the samples, and an alachlor fortified control for each set of 3 strips. Remove the number of well strips needed (twelve wells per strip) . Refrigerate the unused strips in the resealable plastic bag containing a desiccant provided in the kit. ”