Skip to main content
. Author manuscript; available in PMC: 2012 Apr 15.
Published in final edited form as: Dev Biol. 2011 Jan 22;352(2):215–227. doi: 10.1016/j.ydbio.2011.01.010

Table 1.

Experimental details for immunohistochemistry experiements

Immunohistochemistry experiments with BrdU
Experiment Citric acid boil CH3OH:H202 Block Primary Antibody Secondary antibody and detection Biotin block Other Primary Antibody Other secondary and detection
BrdU and LH 5 min no TNB1 1:100 anti-BrdU, Oxford Biotech anti-rat FITC no 1:500 anti-LH, NHPP2 anti-guinea pig biotin, Streptavidin Cy3
BrdU and TSH 5 min yes TNB 1:100 anti-BrdU, Ab Serotec anti-rat biotin, Streptavidin Cy3 yes3 1:1000 anti-TSH, NHPP anti-rabbit biotin, TSA FITC
BrdU and POMC 5 min yes TNB 1:100 anti-BrdU, Ab Serotec anti-rat biotin, Streptavidin Cy3 yes 1:1500 anti-POMC, NHPP anti-rabbit biotin, TSA FITC
BrdU and GH 5 min yes TNB 1:100 anti-BrdU, Ab Serotec anti-rat biotin, Streptavidin Cy3 yes 1:1000 anti-GH, NHPP anti-human biotin, TSA FITC
BrdU and all hormones 5 min no TNB 1:100 anti-BrdU, Ab Serotec anti-rat biotin, Streptavidin Cy3 yes same as above, combined anti-guinea pig, rabbit, human, Streptavidin Cy3
BrdU and Sox2 10 min yes TNB 1:100 anti-BrdU, Ab Serotec anti-rat biotin, Streptavidin Cy3 yes 1:2000 anti-Sox2, Chemicon anti-rabbit biotin, TSA FITC
Immunohistochemistry experiments without BrdU
TBX19 and POMC 5 min no TNB 1:1500 anti-POMC, NHPP anti-rabbit biotin, Streptavidin-488 yes 1:200 anti-TBX19, gift from J. Drouin anti-rabbit biotin, Streptavidin Cy3
NR5A and LH 5 min no TNB 1:1000 anti-NR5A, gift from G. Hammer anti-rabbit biotin, Streptavidin-488 yes 1:500 anti-LH anti-guinea pig biotin, Streptavidin Cy3
POU1F1 and GH, TSH 5 min yes TNB 1:600 anti-POU1F1, gift from S. Rhodes anti-rabbit biotin, Streptavidin-488 yes 1:100 anti-GH, 1:100 anti-TSH anti-rabbit and anti-human biotin, TSA TRITC
pSMAD1/5/8 10 min yes GBT4 1:100 anti-pSMAD, Upstate Biological anti-rabbit biotin, TSA FITC no
pMAPK 10 min yes GBT 1:100 anti-pMAPK, Cell Signaling anti-rabbit biotin, TSA FITC no
1

Block from Perkin-Elmer TSA-FITC Kit

2

National Hormone and Pituitary Program

3

Avidin/Biotin Blocking Kit from Vector Laboratories

4

10% Goat Serum, 1% BSA, 0.1% Triton X-100