I am going to check the PGE2 secretion by using Prostaglandin E2 Express EIA KIT, cayman chemical. I am using it for the first time, so little hesitated. I have four samples, as below:
(1) Control (2) PGE2 treated only (3) Test Drug treated (4) PGE2 pre-treated, followed by drug treated.
They have provided booklet with the KIT, given in the link below:
https://www.caymanchem.com/pdfs/500141.pdf
I am working with Adherent cells, and i have to take Cells supernatant as my sample.
They have mentioned five types of wells.
(1) Blank (2) TA (3) NSB (4) B0 (5) Std/sample
So does it mean that from each of my samples [(1) Control (2) PGE2 treated only (3) Test Drug treated (4) PGE2 pre-treated, followed by drug treated.], i have to fill these five wells, along with the prescribed reagents of the KIT?
Hi Salam,
regarding your question ".. that from each of my samples ... fill these five wells, ...":
No, once only you have to fill these five "control" wells for all your samples ( "one for all - and all for one", as the muketeer coined ;-)... );
these five well are recomanded by cayman (as they are recommanded by other supplier) to ensure the recording of any assay / sample preparation related influence;
depending on your research question you can / have to consider the respective measured values:
a) (1) Blank and/or (3) NSB) subtration from values of standards (as explained in the cyaman booklet)
b) (2) TA and (4) Bo for calculation the standard curve (as explaned in the cayman booklet)
c) considering (3) NSB by calculation/interpretation of your real sample (1), (2), (3), and (4) - > whether or not you really revealed an effect by treating you samples.
wish good success by sample preparation, assay procedure, and - intrepretation of your experimental results.
best regards
Dirk
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