IUBio

Lowry protein determination

Kyle Legate legatek at sympatico.ca
Wed Jun 25 10:41:50 EST 2003


Charter News wrote:
>
> I have a problem to construct a standard curve by using Bovine Serum
> Albumin. The standard curve is linear, but it intercepts y-axis at
> 0.248 Abs (Beckman spectrometer). The sample values I want to compare
> with the standard curve are lower than the y-intercept value, e.g.
> 0.0987. Is that the problem with the BSA? Or there is something wrong
> with my sample readings?
>
Do you mean that your blank with no protein gives a reading of 0.248? When I
make a standard curve I always blank the instrument with a sample containing
no protein (try to keep buffer components constant between your standadr and
your sample to eliminate the effect of these components on the colour
development); this ensures that the standard curve goes through 0. In any
case it doesn't sound like you have much protein in your sample. Perhaps try
a more sensitive technique (BCA assay microtitre plate method?)





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