Table 1 Antibodies used in the present study, source, dilution, type, species reactivity, immunogen, negative and positive controls used.
Antibody/Source | Dilution/antibody type | Species reactivity/immunogen *** | Negative control | Positive control/cellular location |
|---|---|---|---|---|
Pan neurofilament/Zymed | 1:500/mouse monoclonal | Rat, mouse, human/human cocktail neurofilament low, middle and high molecular weight peptides | No antibody on ICC | Mouse, rat human, cerebellar cortex*/neuron perikarya and axons |
Synapsin-1/Chemicon | 1:1000/rabbit polyclonal | Rat, bovine, human/synapsin I (Ia and Ib) bovine brain | No antibody on ICC | Mouse, rat cerebellar cortex*/nerve terminals |
Calbindin/Sigma | 1:1000/mouse monoclonal | Rat, mouse, human, goat/bovine kidney – calbindin D-28k | No antibody on ICC | Mouse, rat cerebellar cortex*/Purkinje cell arborization |
Parvalbumin/Chemicon | 1:1000/mouse monoclonal | Rat, human, bovine/Parvalbumin purified from frog muscle | No antibody on ICC | Purkinje cell arborization, basket cells mice and human cerebella |
SOD-1/Santa Cruz | 1:500/rabbit polyclonal | Rat, mouse, human/aa 1–154 full length human SOD-1 | Antibody absorption with peptide on ICC | Rat* and mice** intestine/luminal epithelial cells |
SOD2/Abcam | 1:1000/rabbit polyclonal | Drosophila and several vertebrates/full length rat SOD-2 protein | No antibody on ICC | Mouse and rat cerebellar cortex and brain stem/neuronal soma |
HO-1/Santa Cruz | 1:500/Goat polyclonal | Rat, mouse, human/peptide mapping C-terminus of HO-1 of rat origin | Antibody absorption with peptide on ICC | Rat postnatal day 6 cerebellum cortex*/neurons and blood vessels |
HO-2/ | 1:1000/Rabbit | Rat, mouse, human, rabbit, cow, dog/full length protein | No antibody on ICC | Rat cerebellum*/Purkinje cell soma |
Abcam iNOS/Chemicon | polyclonal 1:1000/rabbit polyclonal | Rat, mouse/C-terminal 14 aa peptide from mouse macrophage iNOS | No antibody on ICC | LPS mouse brain, inner ear*/macrophages in blood vessels |
nNOS/Calbiochem | 1:1000/rabbit polyclonal | Rat, mouse, human/peptide corresponding amino acids 724–739 of human brain NOS coupled to KHL | No antibody on ICC | Normal rat, mice, PCs and layer 5 primary neurons |
eNOS/Chemicon | 1:500/mouse monoclonal | Rat, mouse, bovine, human/Bovine eNOS purified from cultured aortic EC | No antibody on ICC | Blood vessels from human and mouse brain. |
GFAP/Sigma | 1:1000/Mouse monoclonal | Rat, pig, human/Purified GFAP from pig spinal cord | No antibody on ICC | Human cortex and cerebella. |
Nitrotyrosine/Chemicon | 1:1000/mouse polyclonal | Rat, mouse, human/nitrated KLH | No antibody on ICC | Normal human cerebellum** 95 year old/neuronal somata |
GAD67/Chemicon | 1:2000/mouse monoclonal | Rat, mouse, human/Recombinant GAD67 protein | No antibody on ICC | Normal mouse brain and cerebellum. |
Ferritin H/Santa Cruz | 1:500/rabbit polyclonal | Rat, mouse, human/aa 131–183 of ferritin heavy chain of human | No antibody on ICC | Normal human cerebellar cortex** 98 year old/neuronal somata |
Ferritin L/Sigma | 1:1000/rabbit polyclonal | Rat, mouse, human horse spleen ferritin | No antibody on ICC | Normal human cerebellar cortex** 98 year old/glial cells |