RE: negative controls

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From:"Tarpley, John" <> (by way of histonet)
To:histonet <>
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You want to use the negative serum at the same concentration as your primary
which is not necessarily the same dilution. The data sheets for both the
primary and the control should give you an Ig concentration in probably
micrograms/ml. If you don't have that information you'll need to call the
vendor's).  Since you know the dilution of your primary you can calculate
the working concentration of your primaryby dividing the stock concentration
by the dilution you use. For the negative control you can then divide the
negative serum stock concentration by the working primary ab concentration
to give the dilution for the negative serum. Hope this helps.

John Tarpley 15-2-B
Specialist Image Analysis & Immunohistochemistry
Amgen Inc
One Amgen Center Drive
Thousand Oaks, CA  91320

Views expressed are mine alone and do not represent the views of my employer

> ----------
	--Snip Snip--  All of the buzz a few weeks ago convinced me that it
was a good
> idea to use sera from the animal the primary was made in.  Using the
> control antibody at the same dilution as my primary I am now seeing
> staining.  However, I see only what I would expect to see labelled with my
> primary Ab.  Does this mean that I have to futz around with my conditions
> until I see no labelling using my negative control antibody even though I
> see a total different staining pattern than that of my primary?
> Thanks again!!!!
> Nora
> Nora Fox
> Division of Orthopaedic Surgery
> Stanford University
> Edwards R144, MC 5341
> 300 Pasteur Drive
> Stanford, CA 94305
> phone: (650)725-2746
> fax:  (650)723-6396

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