Table 1 List of antibodies used in the study
Antibody/dye | Specificity | Immunogen/clone | Dilution | Source | Catalog# |
|---|---|---|---|---|---|
Primary antibodies for IHC | |||||
Rabbit anti ionized calcium-binding adaptor molecula 1 (Iba-1) | 17 kDa EF hand protein expressed on macrophages and microglia | Synthetic peptide of C-terminus Iba-1 N’-PTGPPAKKAISELP-C’ | 1:1000 | Wako | 019-19741 |
Rat anti mouse CD86 | Mouse B7.2 | LPS-activated CBA/Ca mouse splenic B cells | 1:600 | Biolegend | 105010 |
Rabbit anti protein gene Product 9.5 (PGP9.5) | All mammalian species, PGP9.5 expressed on neuronal cell bodies and axons in central and peripheral neural system | Human PGP 9.5 Protein purified from pathogen-free human brain | 1:800 | Ultraclone | RA95101 |
Rabbit anti human activating transcription Factor 3 (ATF3) | Mouse, rat and human, ATF3 expressed on damaged neurons | A peptide mapping at the C-terminus of human ATF-3 | 1:2000 | Santa cruz | SC188 |
Rat anti mouse CD8 alpha | Mouse CD8 alpha | Mouse thymus/spleen cells | 1:2000 (fresh frozen tissues with short fixation) | BD pharmingen | 550281 |
Rabbit anti glucose transporters-1 (Glut1) | Mouse, rat and human, Glut1expressed on endothelial cells and perinurium | Synthetic peptide corresponding amino acids with the C-terminus of human GLUT-1 coupled to KLH (C-ELFHPLGADSQV) | 1:6000 | Merck millipore | 07-1401 |
Secondary antibodies for IHC | |||||
Alexa fluor 594 goat anti-rabbit IgG | IgG heavy chains and all classes of IgG light chains from rabbit | 1:500 | Molecular probes (Invitrogen) | A11037 | |
Alexa fluor 488 goat anti-rabbit IgG | IgG heavy chains and all classes of IgG light chains from rabbit | 1:500 | Molecular probes (Invitrogen) | A11034 | |
Alexa fluor 594 rabbit anti-rat IgG | IgG heavy chains and all classes of IgG light chains from rat | 1:500 | Molecular probes (Invitrogen) | A11007 | |
Alexa fluor 488 rabbit anti-rat IgG | IgG heavy chains and all classes of IgG light chains from rat | 1:500 | Molecular probes (Invitrogen) | A11006 | |
Dye | |||||
4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) | 1:15,000 | Sigma-aldrch | D9542 | ||
Fluoro myelin red fluorescent myelin stain | 1:300 | Molecular probes (Invitrogen) | F34652 | ||
Flow cytometric antibodies | |||||
FITC rat anti-mouse CD11b | Mouse CD11b+ cells | C57BL/10 splenic T cells and concanavalin A-activated C57BL/10 splenocytes | 1:50 | BD pharmingen | 553310 |
Rat anti-mouse CD45 APC | Mouse CD45+ cells | Clone 30‒F11 | 1:50 | eBioscience | 17-0451-82 |
Rat anti-mouse CD8 alpha PerCP-Cy5.5 | Mouse CD8+ cells | Clone 53-6.7 | 1:50 | eBioscience | 45-0081-82 |
Rat anti-mouse CD86 (B7.2) PE | Mouse B7.2+ cells | Clone GL1 | 1:50 | eBioscience | 12-0862-82 |