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Table 1 Primary antibodies and modes of antigen retrieval for immunohistochemistry

From: Microglia pre-activation and neurodegeneration precipitate neuroinflammation without exacerbating tissue injury in experimental autoimmune encephalomyelitis

Antibody Origin Dilution Antigen retrieval CSA Source
Amyloid precursor protein (APP) [clone 22C11] Mouse (mAb) 1:1000 St (C)   Chemicon, Merck Millipore; MAB348
CD3 [clone SP7] Rabbit (mAb) 1:2000 St (E) yes Neomarkers; RM-9107
CD68 [clone ED1] Mouse 1:10,000 St (E)   Serotec
2′,3’-Cyclic-nucleotide 3′-phosphodiesterase (CNPase) [clone SMI 91] Rabbit (mAb) 1:2000 St (E)   Sternberger Monoclonals; SMI91
Glial fibrillary acidic protein (GFAP) Rabbit (pAb) 1:3000 St (E)   Dako; Z0334
Ionized calcium-binding adapter molecule 1 (Iba-1) Rabbit (pAb) 1:3000 St (E)   Wako Chemicals; 019–19741
Inducible nitric oxide synthase (iNOS) Rabbit (pAb) 1:750 St (C)   Chemicon, Merck Millipore; AB16311
MHC class II RT1B [clone OX-6] Mouse (mAb) 1:250 St (E)   Serotec; MCA46G
Neuronal nuclei (NeuN) [clone A60] Mouse (mAb) 1:250 St (C)   Chemicon, Merck Millipore; MAB377
Chondroitin sulfate proteoglycan 4 (NG2) Rabbit (pAb) 1:250 St (E)   Chemicon, Merck Millipore; AB5320
Oligodendrocyte transcription factor 2 (Olig2) Rabbit (pAb) 1:1000 St (E)   Chemicon, Merck Millipore; AB9610
p22phox [clone FL-195] Rabbit (pAb) 1:100 for pre-absorptiona St (C)   Santa Cruz Biotechnology; sc20781
Purinergic receptor P2Y12 (P2RY12) Rabbit (pAb) 1:1000 St (E) yes Kindly provided by Oleg Butovsky (Harvard Medical School)
Transmembrane protein 119 (TMEM119) Rabbit (pAb) 1:1000 St (E)   Synaptic Systems; 400002
ATP synthase alpha [clone 7H10] Mouse (mAb) 1:200 St (E)   Thermo Fisher Scientific; 459240
Phosphorylated neurofilament H [clone SMI 31] Mouse (mAb) 1:20,000 St (E)   Affiniti; NA1219
  1. St, steaming of sections using the indicated buffer solution; C, 10 mM citrate buffer (pH 6.0); E, 1 mM EDTA in 10 mM Tris (pH 8.6); mAb, monoclonal antibody; pAb, polyclonal antibody; CSA, catalysed signal amplification via biotinylated tyramine enhancement
  2. ap22phox antibody was pre-absorbed with Lewis cortex homogenate in order to reduce unspecific background staining