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. 2015 Aug 21;29(10):1510–1521. doi: 10.1210/me.2015-1192

Table 1.

Immunohistochemistry Experiments

Experiment Citric Acid Boil CH3OH: H2O2 Block Primary Antibody Secondary Antibody and Detection Biotin Block Other Primary Antibody Other Secondary and Detection
ISL1 10 min-embryo 15 min-postnatal Yes M.O.M.,a TNBb 1:600 anti-ISL1, DSHBc Antirat biotin, TSA FITC
ISL1 and POU1F1g 10 min-embryo 15 min-postnatal Yes M.O.M., TNB 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC Yes 1:500 anti-POU1F1, gift from S. Rhodes Antirabbit biotin, TSA TRITC
ISL1 and NR5A1g 10 min Yes M.O.M., TNB 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC Yes 1:1000 anti-NR5A, gift from G. Hammer Antirabbit biotin, TSA TRITC
ISL1 and TBX19g 10 min Yes M.O.M., TNB 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC Yes 1:200 anti-TBX19, gift from J. Drouin Antirabbit biotin, TSA TRITC
CGA and ISL1 10 min Yesf M.O.M., TNB 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC Yes 1:1000, anti-CGA, NHPPd Antirabbit biotin, TSA TRITC
TSHb and ISL1 15 mine Yesf M.O.M., TNB 1:1000 anti-TSHb, NHPP Antirabbit biotin, TSA TRITC Yes 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC
LHb and ISL1 15 mine Yesf M.O.M., TNB 1:500 anti-LHb, NHPP Antiguinea pig, TSA TRITC Yes 1:600 anti-ISL1, DSHB Antirat biotin, TSA FITC
GH none Yes NGSh 1:1000 anti-GH,j NHPP Antihuman biotin, SA Cy2i
TSH none Yes NGS 1:1000, anti-TSH, NHPP Antirabbit biotin, SA Cy2i
TSH and POU1F1 10 mine Yesf NGS M.O.M. 1:1000, anti-TSH, NHPP Antiguinea pig, TSA TRITC Yes 1:500, anti-POU1F1 Antirabbit biotin, TSA FITC
TSH and NR5A1 10 mine Yesf NGS M.O.M. 1:1000, anti-TSH, NHPP Antiguinea pig, TSA TRITC Yes 1:1000, anti-NR5A1 Antirabbit biotin, TSA FITC
a

Mouse kit from Vector Labs.

b

Block from PerkinElmer Tyramide Signal Amplification TSA Fluorescein isothiocyanate FITC and Tetramethylrhodamine (TRITC) kit.

c

Developmental Studies Hybridoma Bank.

d

National Hormone and Pituitary Program.

e

Citrate boiling done after primary antibody.

f

Second CH2OH:H2O2 block done before second primary antibody.

g

Primary antibodies incubated at same time.

h

5% normal goat serum, 3% BSA in 1× PBS.

i

Streptavidin cyanine 2.

j

Anti-GH antibody left on for 1 hour.