Fernando,
Including the detergent into the standard
solutions reduces the errors.
If this doesn't work with your system, you could precipitate the
proteins in your sample (e.g. with TCA or try a "reverse"
phenol/chloroform extraction and keep the pellet) and wash out the
detergent.
Wolfgang
> From: "Dr. Fernando Rodriguez-Pascual" <frodrigu at mail.uni-mainz.de>
> Subject: Nonidet P40 and Bradford
> Date: Mon, 10 Jan 2000 15:13:24 +0200
> Organization: Johannes Gutenberg-Universitaet Mainz, Germany
> Reply-to: frodrigu at mail.uni-mainz.de> To: methods at hgmp.mrc.ac.uk
> Hi all,
>> Have any of you experienced problems in performing protein
> determinations by Bradford using extracts containing 0.5% Nonidet
> P40. I mean, whether it is compatible with the Bradford reagent and
> up to which concentration. In former times I used to have no
> problems for that but now I find that anyhow it affects the assay.
> By the way, Sigma is not longer purchasing Nonidet P40, but a
> "chemically indistinguible from it" compound (as Sigma) called
> IGEPAL-CA 630. Do you know if it is really the same? Have you found
> problems by using this new nonionic detergent?
>> Chao
>> Fernando
> --
> #############################################
> Dr. Fernando Rodríguez Pascual
> Department of Pharmacology
> Johannes Gutenberg University
> Obere Zahlbacher Str. 67
> D-55101 Mainz, Germany
> Tf. +49 (6131) 39 3 68 36 or 39 3 32 45
> Fax. +49 (6131) 39 3 66 11
> email: frodrigu at mail.uni-mainz.de> #############################################
Dr. Wolfgang Schechinger, Pathobiochemistry Dept.
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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