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DMU Acetyl-Histone and c-Fos Immuno (view source)
Revision as of 13:00, 3 March 2022
, 13:00, 3 March 2022added dilutions
'''Rats arrive''' (2022-02-23)
'''Rats arrive''' (2022-02-23)
male (n=8) DOB: 2021-12-27
male (n=8) DOB: 2021-12-27
female (n=8)DOB: 2021-12-6,
female (n=8) DOB: 2021-12-6
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'''Surgery'''
'''Groups'''
4 groups for each sex: NaCl-NaCl, LiCl-NaCl, NaCl-NaB, LiCl-NaB
4 groups for each sex: NaCl-NaCl, LiCl-NaCl, NaCl-NaB, LiCl-NaB
'''Immunohistochemistry'''
'''Immunohistochemistry'''
Usual IHC protocol: 30 min Triton, overnight primary, 1 h secondary, mount with VectaShield w/DAPI.
anti-acetyl-Histone Primary: anti-acetyl-Histone H3 (Lys14) rabbit polyclonal, Millipore 07-353, Lot #3259085, r. 2019-09-26 (use at 1:2000)
anti-acetyl-Histone Primary: anti-acetyl-Histone H3 (Lys14) rabbit polyclonal, Millipore 07-353, Lot #3259085, r. 2019-09-26 (use at 1:2000)
anti-c-Fos Primary: anti-c-Fos monoclonal antibody (E-8), Santa Cruz sc-166940, Lot # L1621, r. 2022-3-3 (use at 1:100)
anti-c-Fos Primary: anti-c-Fos monoclonal antibody (E-8), Santa Cruz sc-166940, Lot # L1621, r. 2022-3-3 (use at 1:100)
Secondary: Texas Red conjugated Goat Anti-Mouse, Jackson Labs 115-075-146, Lot #72021, r. 2008-05-24 (use at 1:100)
Secondary: Texas Red conjugated Goat Anti-Mouse, Jackson Labs 115-075-146, Lot #72021, r. 2008-05-24 (use at 1:100)
2022-3-3 thru 2022-3-4: DMU01-DMU16 hindbrain sections processed (12 sections each, 500 λ/well).
Dilutions
16 wells x 500 λ = 8000 λ, so make 9000 λ
Primaries:
9000 λ PBS/BSA
anti-acetyl-Histone at 1:2000 => 9000/2000 = 4.5 λ
anti-c-Fos at 1:100 => 9000/100 = 90 λ
Secondaries:
9000 λ PBS/BSA
anti-Rabbit at 1:100 => 9000/100 = 90 λ
anti-Mouse at 1:100 => 9000/100 = 90 λ
[[Category:Taste Aversion]]
[[Category:Taste Aversion]]
[[Category: CTA Experiments]]
[[Category: CTA Experiments]]