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Dear Sir, Dear Madame,I tried to use your protocol for GAGs...
MB
Matteo Beretta Jun 22, 2017
Dear Sir, Dear Madame,
I tried to use your protocol for GAGs quantification in cell colture medium conditioned by intervertebral disc cells for 24h-48h and used the bovine chondroitin 4-sulfate as standard. I am having huge problems even with the standard curve (a eight point-cu Read more
I tried to use your protocol for GAGs quantification in cell colture medium conditioned by intervertebral disc cells for 24h-48h and used the bovine chondroitin 4-sulfate as standard. I am having huge problems even with the standard curve (a eight point-cu Read more
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Matteo Beretta Answered Jun 23, 2017
Fondazione Ca' Granda - Ospedale Maggiore Policlinico
I want to thank you very much for your quick answer!
So do you think that the problem is the L-Cysteine and not the FBS? In light of your scientific experience could you briefly explain me why L-Cysteine could be a problem for the GAGs quantification with the DMMB solution?
Thank you very much again for your help! It's very much appreciated!
Kind regards,
MB
So do you think that the problem is the L-Cysteine and not the FBS? In light of your scientific experience could you briefly explain me why L-Cysteine could be a problem for the GAGs quantification with the DMMB solution?
Thank you very much again for your help! It's very much appreciated!
Kind regards,
MB
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0 comments ShareTarsis Ferreira Author Answered Jun 22, 2017
University of Houston
Assuming you media contains L-Cysteine, add EDTA to 0.1 mM and Papain, 0.5 U/mg (Sigma, Cat. No. 76216 - 25 U/ml). Digest samples in papain solution for 15–18 hours at 65°C. Digested samples can be stored at −20°C.
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