24
edits
Changes
→NEFA determination from serum (use Wako Diagnostics NEFA-HR(2) Microtiter Kit)
#To wells of a clear 96 well plate, add 4uL of water (as blank), appropriate standard (the kit comes with stock of 1 mEq/L. Dilute the stock in ddH20 to give 0.25 and 0.5 mEq/L standards. For the high standard, add 8 uL of the 1 mEq/L stock. Due to the extra volume, the concentration of this last standard is actually 1.97 mEq/L).
#To remaining wells, add 4 uL of serum samples. ***Note: You may need to add more/less serum, depending on the NEFA concentration of your samples. The assay is linear between 0.01-4.00 mEq/L***
#To each well, add 225 uL of reaction buffer A, mix gently and incubate at 37 deg C for ~5 minutes.
#Allow plate to return to room temperature before reading at 560:670 nm. These are the initial readings.
