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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Med Primatol. 2014 Aug 1;43(6):468–476. doi: 10.1111/jmp.12138

Table 2.

Antibody source, dilution, and antigen retrieval for immunohistochemistry protocols.

Marker Epitope
Retrieval		 Primary
Antibody		 Incubation
Time		 Dilution Secondary
Antibody		 Dilution
(all 20
min)		 Tertiary
Antibody
(all 30 min)
GFAP None PAH1, Dako
(Carpinteria, CA)		 45 min, room
temperature		 1:340 GAR-b2 1:200 Vectastain
ABC Elite3
Vimentin Proteinase K
(5 min, RT)		 MAH4, Clone 3B4,
Dako		 Overnight,
4°C		 1:162 HAM-b5 1:200 Vectastain
ABC
Standard3
Glut-1 Microwave6 PAH1, Millipore
(Billerica, MA)		 30 min, room
temperature		 1:1000 GAR-b2 1:200 Vectastain
ABC
Standard3
CNPase Microwave6 MAH4, Clone 11-
5B, NeoMarkers
(Fremont, CA)		 60 min, room
temperature		 1:200 HAM-b5 1:200 Vectastain
ABC Elite3
S100 Microwave6 PAH1, S100, Dako 60 min, room
temperature		 1:3200 GAR-b2 1:200 Vectastain
ABC Elite3
Iba1 Microwave6 PAH1, Wako
(Richmond, VA)		 30 min, room
temperature		 1:1000 GAR-b2 1:200 Vectastain
ABC Elite3
Ki67 Microwave6 MAH4, Clone MIB-
1, Dako		 Overnight,
4°C		 1:35 HAM-b5 1:200 Vectastain
ABC Elite3
p53 Microwave6 MAH4, Clone DO-
7, AbD Serotec
(Raleigh, NC)		 Overnight,
4°C		 1:500 Dual Link
System,
Dako		 None None
SIV Microwave6 MAH4, HIV-1 p247 Overnight,
4°C		 1:400 HAM-b5 1:200 Vectastain
ABC Elite3
SV40 Microwave6 MAH4, SV40 large
T antigen, Clone
Pab416,
Oncogene
(Billerica, MA)		 Overnight,
4°C		 1:25600 HAM-b5 1:200 Vectastain
ABC Elite3
CMV Microwave6 serum, CMV, IEI
(exon4), PA
Barry, UCD-CCM
(Davis, CA)		 30 min, room
temperature		 1:1600 GAR-b2 1:200 Vectastain
ABC
Standard3
LCV Microwave6 MAH4, EBNA2,
Clone PE2, Leica
(Buffalo Grove, IL)		 Overnight,
4°C		 1:600 HAM-b5 1:200 Vectastain
ABC Elite3
1

Polyclonal anti-human

2

Biotinylated goat anti-rabbit, Vector Laboratories, (Burlingame, CA, USA)

3

Vector Laboratories

4

Monoclonal anti-human

5

Biotinylated horse anti-mouse, Vector Laboratories

6

Microwave in sodium citrate buffer for 20 minutes, followed by cooling at room temperature for 20 minutes

7

The following reagent was obtained through the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 p24 Monoclonal Antibody (183-H12-5C) from Dr. Bruce Chesebro and Kathy Wehrly.