RE: [Histonet] mixing up primary and secondary antibodies fordouble/triple staining

From:"Elizabeth Chlipala"


You can combine the primaries if all of them are from different species.
If they are from the same species you will have to perform the staining
sequentially.  We have some images posted on our web page of
immunofluorescent staining on 50 micron vibratome sections that have
been done by combining the primary and secondary reagents.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
[] On Behalf Of Yves
Sent: Tuesday, July 19, 2005 11:14 AM
Subject: [Histonet] mixing up primary and secondary antibodies
fordouble/triple staining

Dear Histonetters,
I have a general question on double or triple fluorescence stainings. 
Jackson Immunoresearch recommends doing sequential stainings, i.e. 
complete staining for antigen A first, then antigen B and then antigen 
C. Is there any danger in mixing up all the primary antibodies in one 
mix instead of using the sequential approach ? And, if using secondary 
antibodies derived from the same species, is there any danger in doing 
the same for the secondaries ?

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