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JG Selenate and LiCl-induced pMAPK and c-Fos (view source)
Revision as of 12:58, 12 January 2024
, 12:58, 12 January 2024created page
Expt Code: JG
TODO: get dates and number of rats, DOBs
IACAUC Protocol: 1113
# Animals
* Adult male Sprague Dawley rats (n=12)
* weight 300 - 500g
* Source: Charles River
* DOB:
# Procedure (rev 2013-7-24)
Weight rats
## Schedule:
(Stagger pairs of rats by 20 min to allow time for perfusion)
at 0h, inject with either 1 ml/kg vehicle (0.15 M NaCl) or 0.5 mg/kg Na2SeO4
at 1 h, inject with either 12 ml/kg NaCl or 12 ml/kg LiCl (0.15 M)
at 2 h, overdose with Sleepaway and perfuse
Dissect brains, postfix for 2 h in 4%PFA, rinse with 0.1 M PB, cyroprotect in 30% sucrose
# Immunohistochemistry
Process brains for immunohistochemistry with anti-cFos and anti-pMAPK:
From Kwon and Houpt:
anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) rabbit mAb (#4370, 1:500, Cell Signaling Technology)
From Kimbrough TORC-1 paper:
anti-c-Fos goat polyclonal antibodies (#SC-52-G, 1:10,000 Santa Cruz Biotechnology)
TODO: get dates and number of rats, DOBs
IACAUC Protocol: 1113
# Animals
* Adult male Sprague Dawley rats (n=12)
* weight 300 - 500g
* Source: Charles River
* DOB:
# Procedure (rev 2013-7-24)
Weight rats
## Schedule:
(Stagger pairs of rats by 20 min to allow time for perfusion)
at 0h, inject with either 1 ml/kg vehicle (0.15 M NaCl) or 0.5 mg/kg Na2SeO4
at 1 h, inject with either 12 ml/kg NaCl or 12 ml/kg LiCl (0.15 M)
at 2 h, overdose with Sleepaway and perfuse
Dissect brains, postfix for 2 h in 4%PFA, rinse with 0.1 M PB, cyroprotect in 30% sucrose
# Immunohistochemistry
Process brains for immunohistochemistry with anti-cFos and anti-pMAPK:
From Kwon and Houpt:
anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) rabbit mAb (#4370, 1:500, Cell Signaling Technology)
From Kimbrough TORC-1 paper:
anti-c-Fos goat polyclonal antibodies (#SC-52-G, 1:10,000 Santa Cruz Biotechnology)