[Histonet] RE: Normal donkey serum block solution

[C.M. van der Loos]

   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