Antibodies used in immunocytochemistry
| # | Antibody | Origin | Target | Dilution | Antigen retrieval | Source |
|---|---|---|---|---|---|---|
| 1 | PLP | Mouse (mAB) | PLP | 1:1000 | St (E) | MCA839G; Serotec |
| 2 | CD68 | Mouse (mAB) | Phagocytic macrophages | 1:100 | St (E) | M0814; Dako |
| 3 | NF | Rabbit (pAB) | NF 150 kDa | 1:2000 | St (E) | AB1981; Chemicon |
| 4 | 8OHdG | Goat (pAB) | 8-Hydroxy 2-deoxy guanosine | 1:1000 | St (E)* | Abcam, ab10802 |
| 5 | APP | Mouse (mAB) | APP | 1:1000 | St (C) | MAB348; Chemicon |
| 6 | CD3 | Rabbit (mAB) | T cells | 1:2000 | St (E)† | RM-9107-S; Neomarkers |
| 7 | E06 | Mouse (mAB) | Oxidised phospholipids | 10 µ/mL | 0 or St (C or E) | Palinski et al, 199643 |
| 8 | IBA-1 | Rabbit (pAB) | IBA-1 | 1:3000 | St (E)* | 019-19741; WAKO Chemicals |
| 9 | iNOS | Rabbit (pAB) | iNOS | 1:30000 | St (E) | AB5384; Chemicon |
| 10 | MBP | Rabbit (pAB) | Myelin basic protein | 1:2500 | 0 | A0623; Dako |
| 11 | P22phox | Rabbit (pAB) | NADPH oxidase protein | 1:100 | St (C) | sc-20781; Santa Cruz |
| 12 | TPPP/p25 | Rabbit (pAB) | Oligodendrocytes | 1:250 | St (E)* | G. G. Kovacs, Vienna |
| 13 | GFAP | Rabbit (pAB) | Astrocytic GFAP | 1:2000 | St (E)* | Z0334; Dako |
| 14 | MAP-2 | Mouse (mAB) | Neurons | 1:100 | St (E)* | M4403; Sigma |
This table lists antibodies that were used in this study. Antibodies #1–3 were used for staining in all cases of multiple sclerosis (MS) and all controls. Antibodies #4–11 were used for staining in a subsample of 31 MS cases displaying deep grey matter and in all controls. Antibodies #12–14 and also antibody #8 were used for double-labelling with iron.
*Antibody labelling visualised with Fast blue B instead of routinely used 3,3′-diaminobenzidine.
†3,3′-Diaminobenzidine development enhanced by biotinylated tyramine amplification.
0, no antigen retrieval; APP, amyloid precursor protein; C, citrate buffer (pH 6.0); E, EDTA buffer (pH 9.0); GFAP, glial fibrillary acidic protein; IBA-1, ionised calcium-binding adapter molecule 1; iNOS, inducible nitric oxide synthase; mAB, monoclonal antibody; NADPH, nicotinamide adenine dinucleotide phosphate; NF, neurofilament; pAB, polyclonal antibody; PLP, proteolipid protein; St, steaming of sections using the indicated buffer solution.