[Histonet] Oct-3/4

[Richard Cartun]

This is what we do:

Antibody name:  Oct 3/4 (C-20)
- Source:  Santa Cruz Biotechnology (Cat #SC 8629)
- Cat #:  SC-8629
- Made in:  Goat
- Pretreatment:  HIER (vegetable steamer) for 20 min. with citrate
buffer pH6 (DAKO)
- Antibody dilution:  1:100 
- Detection System:  Rabbit anti-Goat-HRP; diluted 1:100 (DAKO)
- Chromogen:  DAB+ (Dako)

Staining is performed on a DAKO Autostainer.

Richard




Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> "Elaine Dooley" <DOOLEEO <@t> shands.ufl.edu> 02/20/06 02:51PM >>>
Dear Histonetters,

I have been having some trouble trying to get this antibody to stain
seminomas.  It is  Oct-3/4 a goat polyclonal antibody from Santa Cruz
Biotechnology.  We also purchased their mouse anti-goat biotin
conjugated secondary.
Any quick hints on making this work.  I have tried it on the Ventana
Benchmark (with changing the secondary antibody).  I have tried
overnight incubations and staining entirely by hand. I have tried
Ventana's CC1 and also tried Cell Marque's trilogy for epitope retrial
but have not had any of the desired nuclear staining the antibody is
supposed to show.

Any helpful hints would be greatly appreciated.

Elaine Dooley 
HTL 
Shands Teaching Hospital
Gainesville FL
352-265-0111 ext 7-2117

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