Hello all,
To measure the presence of some peptides and hormones in the blood it is important to add, at time of collection to the serum/plasma samples, inhibitors of proteases.
At what final concentrations is it recommended to add the following inhibitors?
· DPPIV inhibitor (Millipore Corp or Sigma Aldrich Cat# DPP4 or DPP4-010): Stock solution ready to use, 10ul/ml of serum?
https://www.merckmillipore.com/PT/en/product/DPP-IV-Inhibitor,MM_NF-DPP4
· Aprotinin (Sigma A6279): Stock solution at 10mg/ml (5-10 TIU/ml) and use in serum at a final concentration of 0.1 or 1mg/ml?
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Product_Information_Sheet/a6279pis.pdf
· Serine protease inhibitor (Roche Pefabloc Cat. # 11 429 868 001): Reconstitute a solution at 100mg/ml and use in serum at a final concentration of 1mg/ml?
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Roche/Bulletin/1/pefbscrobul.pdf
· Protease Inhibitor cocktail (Sigma Cat# P2714 ): Reconstitute a solution at 100mg/ml and use in serum at a final concentration of 1mg/ml?
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Datasheet/5/p2714dat.pdf
Thank you very much for your help!
Karine
Hi,
I would read these articles to get a better sense of what you need to do. Since many endogenous protease inhibitors already exist in serum/plasma, most proteomic studies don’t require the addition of extra inhibitors, although some proteins may be more susceptible to specific cleavage than others.
https://www.researchgate.net/post/How_to_determine_concentrations_for_protease_inhibitors_in_serum
Article The effect of different protease inhibitors on stability of ...
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/asbmb2005_quant_proteoly.pdf
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