是关于老鼠瘦素的说明书!!内容是关于化验程序.
Assay Procedure
Bring all reagents to room temperature for at least 30 minutes prior to opening.
All standards and samples should be run in duplicate.
1.Refer to the Assay Layout Sheet to determine the number of wells to be used and put any
remaining strips with the desiccant back into the foil pouch and seal the ziploc. Store unused wells at 4!C.
2.Pipet 100 |L of Assay Buffer into the S0 (0 pg/mL Standard) wells.
3.Pipet 100 |L of Standards #1 through #7 into the appropriate wells.
4.Pipet 100 |L of the Samples into the appropriate wells.
5.Tap the plate gently to mix the contents.
6.Seal the plate and incubate for 1 hour at 37!C.
7.Empty the contents of the wells and wash by adding 400 |L of wash solution to every well.
Repeat the wash 6 more times for a total of 7 washes. After the final wash, empty or aspirate
the wells, and firmly tap the plate on a lint-free paper towel to remove any remaining wash buffer.
8.Pipet 100 |L of the prepared Labeled Antibody into each well, except the Blank.
9.Seal the plate and incubate at 37!C for 30 minutes.
10.Empty the contents of the wells and wash by adding 400 |L of wash solution to every well.
Repeat the wash 8 more times for a total of 9 washes. After the final wash, empty or aspirate the wells, and firmly tap the plate on a lint-free paper towel to remove any remaining wash
buffer.
11.Add 100 |L of the TMB Substrate to each well.
12.Incubate for 30 minutes at room temperature in the dark.
13.Add 100 |L of Stop Solution to each well.
14.Blank the plate reader against the Blank wells, read the optical density at 450 nm, preferably
with correction between 570 and 590 nm. If the plate reader is not able to be blanked against the Blank wells, manually subtract the mean optical density of the Blank wells from all
readings.