Anti-NF-H Antibody (A85337)

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.

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.

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.