[Histonet] phorphprylated-STAT3 antibody

Masaaki Kitada masaakikitada2007 <@t> gmail.com
Wed Jul 30 03:25:57 CDT 2008


Dear all,

I've tried to use anti-phosphorylated-STAT3 antibodies in 10um
thick mouse brain cryosections for immunofluorescence with
antigen retrieval but found dirty result or nothing. Does anyone
know good antibodies against pSTAT3? Or, does anyone let me know
the appropriate protocol for immunofluorescence using following
antibodies?


Antibodies
Cell signaling:
tyr705 9145S
  some nuclei (expected)
  overall cellular processes (unexpected)
tyr705 9131S
  nothing detected

BD Biosciences:
Ser727 612542
  nothing detected

Sample:
Mouse brain section
fix with 4% paraformaldehyde in 0.1M PB and post-fix with the
same fixatives for 1~2 ON
10-um-thick cryosection

Antigen retrieval condition:
  prewarm the solution
  incubate for 20 min
  stay for 10 min
  cool down with running water for 10 min

antigen retrieval solution:
5mM Tris-HCl / 2mM EDTA, pH 8.0


Many thanks,

Masaaki Kitada
Tohoku University Graduate School of Medicine



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