Immunohistological analysis of a rat cerebellum section stained with Anti-NF-H Antibody (1:5,000 | red) and Anti-GFAP Antibody (A85419 | 1:5,000 | green). The blue is DAPI staining of nuclear DNA. Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45µM, and free floating sections were stained with the above antibodies. The Anti-NF-H Antibody labels network of axons of different neurons, while the Anti-GFAP Antibody stains astrocytes and other glial cells.
Mixed neuron/glial cultures stained with Anti-NF-L Antibody (A85453 | green) and Anti-NF-H Antibody (red). Anti-NF-H Antibody binds primarily to the phosphorylated axonal forms of NF-H, in contrast to the Anti-NF-L Antibody which stains both axonal and dendritic/perikaryal neurofilaments. The Anti-NF-L Antibody therefore reveals a prominent cell body in green, while the surrounding axonal profiles are orange, since the are bound by both Anti-NF-L Antibody and Anti-NF-H Antibody. Blue is a DNA stain.
Figure 3: Western Blot - Anti-NF-H Antibody (A85337)
Western blot analysis of spinal cord lysates from different species using Anti-NF-H Antibody (1:20,000 | green): [1] protein standard (red), [2] rat, [3] mouse, and [4] cow spinal cord. Strong band at about 200-220kDa corresponds to the phosphorylated from of NF-H. The protein from different species is known to have different SDS-PAGE molecular weights, with large species generally expressing larger proteins. Smaller proteolytic fragments of NF-H are also detected in spinal cord preparations with this antibody.
Figure 4: Western Blot - Anti-NF-H Antibody (A85337)
Lane labelled CBB shows Coomassie Brilliant Blue stained crude extract of rat spinal cord, with the prominent major neurofilament subunit indicated. In rodents, NF-H runs at 200kDa, NF-M at 145kDa and NF-L at 68kDa. The Rab lane shows a blot probed with Anti-NF-H Antibody (A85336). A prominent band at an apparent SDS-PAGE molecular weight of 200kDa corresponds to phosphorylated form of NF-H.
