Immunofluorescent analysis of rat cerebellum section stained with Anti-NF-L Antibody (1:2,000 | green) and co-stained with Anti-FOX3/NeuN Antibody (A85404 | 1:5,000 | red). Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45µM, and free-floating sections were stained with above antibodies. Anti-NF-L Antibody labels perikarya and processes of neuronal cells, particularly strongly the axons of basket cells, while the Anti-FOX3/NeuN antibody stains the nuclei and proximal cytoplasm of neurons.
Figure 2: Western Blot - Anti-NF-L Antibody (A85286)
Western blot analysis of tissue lysates probed with Anti-NF-L Antibody (1:20,000 | green): [1] protein standard (red), [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord and [6] cow spinal cord. Strong bands at ~68kDa corresponds to NF-L proteins which are known to have slightly different apparent SDS-PAGE molecular weights across species boundaries.
View of mixed neuron/glial cultures stained with Anti-NF-L Antibody (red) and Anti-NF-H Antibody (A85336 | green). The NF-L protein is assembled into neurofilaments which are found throughout the axons, dendrites and perikarya of these cells. In contrast the phosphorylated NF-H has a more restricted expression pattern, being found only in developed axonal neurofilaments. Axonal profiles therefore are yellow since the red and green signals are superimposed. In contrast neurofilaments containing only NF-L appear red.
Figure 4: Western Blot - Anti-NF-L Antibody (A85286)
Western blots of whole protein extracts of rat spinal cord (Lane 1), brain stem (Lane 2), cerebellum (Lane 3) and cerebral cortex (Lane 4). Neurofilaments are concentrated in large projection axons and therefore NF-L is a more prominent component of spinal cord than cortical regions.
