anurag sharma wrote:
> Dear All,
> I am new to this protein chemistry lab and I am trying to do this Micro
> Bradford Assay. The problem is inspite of following protocols for making
> the reagent I am not able to get differential absorbance in blanks and
> samples. Can anyone suggest me the right protocol or point to the
> mistakes.
Since we don't know what you have been up to it is difficult to crystal
ball your mistakes.
If you follow the protocoll given by Marion Bradford things should work:
@article{Bra-76,
AUTHOR= {M.M. Bradford},
TITLE= {A Rapid and Sensitive Method for the Quantitation of
Microgram Quantities of Protein Utilizing the Principles of Protein-Dye
Binding},
JOURNAL= {Anal. Biochem.},
VOLUME= {72},
YEAR= {1976},
PAGES= {248-254},
}
Important is a good quality dye (Serva (Munich) has a good reputation)
and the strict avoidance of interfering substances in the sample, most
notably detergents. Did you check the assay with samples of known
protein concentration (e.g. Immunoglobulin, do not use BSA as this gives
unusually high readings in this assay).
