Table 1. Details of the primary antibodies trialled on 250 μm mouse cerebellar sections and summary of the results.
| Primary Antibody | Target | Host | Isotype | Dilution | Manufacturer and product code | Quality of Staining |
|---|---|---|---|---|---|---|
| anti-Neurofilament H 200 kDa | Neurofilament H (200 kDa) | Rabbit | IgG | 1:200 | Millipore (AB5539) | Specific labelling of axons |
| Anti-Myelin basic protein | Myelin basic protein | Mouse | IgG1 | 1:500 | Biolegend (836502) | Specific labelling of myelin |
| anti-NDUFA13 | NDUFA13 | Mouse | IgG2b | 1:100 | Abcam (ab110240) | Specific positive staining of mitochondria |
| Anti-Porin | Porin | Mouse | IgG2b | 1:100 | Abcam (ab14734) | Specific positive staining of mitochondria |
| Anti-complex IV subunit I | COX1 | Mouse | IgG2a | 1:100 | Abcam (ab14705) | Specific positive staining of mitochondria |
| anti-complex IV subunit IV | COX4 | Mouse | IgG2a | 1:100 | Abcam (ab14744) | Specific positive staining of mitochondria |
| anti-SMI-31 | Phosphorylated neurofilament H & M | Mouse | IgG1 | 1:1000 | Biolegend (801601) | Specific labelling of axons |
| anti-SDHA | Succinate dehydrogenase complex, Subunit A (complex II) | Mouse | IgG1 | 1:100 | Abcam (ab14715) | Specific positive staining of mitochondria |
| anti-parvalbumin | Parvalbumin | Mouse | IgG1 | 1:100 | Swant (235) | Specific labelling of Purkinje Cell soma |
| anti-calbindin D-28k | Calbindin | Mouse | IgG1 | 1:100 | Swant (235) | Specific labelling of Purkinje Cell soma |
| Anti-glut-1 | Endothelial cells | Rabbit | IgG | 1:100 | Thermofisher (PA1–21401) | Staining of both arterioles and capillaries |
| Anti-α smooth muscle actin | Smooth muscle cells | Mouse | IgG2a | 1:100 | Dako (M8051) | In formalin-fixed sections, the smooth muscle layer in arterioles are clearly labelled but there is non-specific staining of nuclei. This non-specific staining is not observed in the 4% PFA-fixed and sucrose frozen section. |
The details of primary antibodies that are commonly used in our laboratory and a summary of the results when applied 250 μm thick mouse cerebellar sections that have been passively cleared.