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Assessing isoproterenol-stimulated whole-body lipolysis in vivo

4 bytes removed, 15:44, 10 March 2015
→‎NEFA determination from serum (use Wako Diagnostics NEFA-HR(2) Microtiter Kit)
== NEFA determination from serum (use Wako Diagnostics NEFA-HR(2) Microtiter Kit) ==
***Note: This page has been updated from a previous version, according to an update on the manufacturers website. Our previous method utilized 2.5 uL serum/standards in 100 uL reaction buffer A and 50 uL reaction buffer B, read at 550:660 nm*** 
#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***