Rick Bright wrote:
>> The other part of this problem is I cannot easily quantitate with
> Bradford or Lowry method in this buffer due to either the SDS (Bradford)
> or the BME (Lowry). Is there another way of
> quantitating (maybe after run onto gel) that would be easier to get
> around this problem?
I developed a method of quantitating proteins which is unaffected by
most buffer components (you can assay proteins dissolved in sample
buffer containing SDS, BME, bromophenol blue etc.).
Basically, the samples and standards are spotted onto a grid drawn by
pencil on Whatman 3MM. After staining in Coomassie blue and destaining,
the dye is eluted and read at 590nm. The details are in the following
reference:
Winterbourne, D.J. Chemical assays for proteins. In: Methods in
Molecular Biology. Volume 19. Biomembrane Protocols I. Isolation and
Analysis, edited by Graham, J.M. and Higgins, J.A.Totowa, New
Jersey:Humana Press, 1993,p. 197-202.
--
Dr. David Winterbourne
Department of Surgery
St George's Hospital Medical School, London SW17 0RE, England
Tel: 0181-725-5581 Fax: 0181-725-3594
