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Protein assay problems

The Grouchybeast thegrouchybeast at hotmail.com
Wed Jan 24 13:04:53 EST 2001

Emir wrote:
> Dot blot your protein, same with the standard
> at different concentrations, wash blots and apply amido black solution
> (something like 0.1 g/100ml in 5% acetic acid, if I am not mistaken). The
> visual comparison of the density of color allows you to estimate
> concentration of the unknown protein. I guess you could develop such an
> assay yourself without buying anything.

No, the kit specifies that the *width* of the dot is used to determine
the concentration, not the colour.  If it had been colour I would have
assumed amido black too. It also claims to be an absolute scale, with no
standards required - the kit comes with a scaler to size the dots on and
get the concentration from.

The protocol is dilute, spot, fix, develop.  Big surprise, there's no
composition given in the on-line documentation for the dilution,
fixative or developing buffers.

I see you standing there, far out along the way,
   I want to touch you but, the night becomes the day.
I count the words that I am never going to say,
   And I see you, in midnight blue.    (ELO - Midnight Blue)

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