[Histonet] RE: Normal donkey serum block solution
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Tue May 3 01:50:23 CDT 2005
Guofeng,
Indeed if you have two secondary antibodies raised in donkey, normal
donkey serum should be used for blocking. Just prepare a
1:10-20 dilution in PBS or TBS and incubate 15 min at RT. Do not wash,
but blot off the normal serum and apply your primary.
In case of two secondaries raised in different species you have to be
aware first there will be no cross-reaction between those species. Two
secondaries from the same species is always the best. However, in the
situation of horse and donkey secondaries, cross-reaction seems highly
unlikely. Just prepare a mixture for blocking composed of normal horse
1:20 and normal donkey 1:20 in PBS or TBS. Otherwise use the
DakoCyto Protein Block, serum-free (X0909).
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone: +31 20 5665631
fax: +31 20 6960389
e-mail: [1]c.m.vanderloos <@t> amc.uva.nl
----- Original Message -----
From
? ? <pex0220 <@t> yahoo.com.cn>
Date
Mon, 02 May 2005 21:30:08 +0800 (CST)
To
histonet <@t> pathology.swmed.edu
Subject
[Histonet] Normal donkey serum block solution
Hello,all,
I am doing double-immunofluorescence. Both the hosts of my secondary
antibodies are donkey, so I plan to use normal donkey serum for
blocking, but I do not know how to make up this solution.
I have ordered donkey serum from sigma company, but the specification
doesnot show how to make up it. so if you have done it, I hope that
you can do me a favor.
In addition, if the hosts of my secondary antiboies are different, one
is horse, the other is donkey, which solution do I use for blocking?
Thank you!
Best wishes!
Guofeng
References
1. mailto:c.m.vanderloos <@t> amc.uva.nl
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