Anti-NF-H Antibody [AH1] (A85340)

Immunofluorescent analysis of rat brain section stained with Anti-Tyrosine Hydroxylase Antibody (A104319), at a dilution of 1:10,000, in red, and co-stained with Anti-NF-H Antibody [AH1] (A85340), at a dilution of 1:1,000, in green. The blue is Hoechst staining of nuclear DNA. 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 the above antibodies. Anti-Tyrosine Hydroxylase Antibody (A104319) stains the striatal TH expressing interneurons, while Anti-NF-H Antibody [AH1] (A85340) labels axons from other neuronal cells.

Western blot detection of the heavily phosphorylated axonal form of NF-H protein (pNF-H) in neural tissue lysates (20µg/lane) with Anti-Neurofilament Heavy Polypeptide Antibody [AH1] (A85340) at dilution of 1:5,000. Lanes on the blot are: [Lane 1] Protein size marker, [Lane 2] Adult rat whole brain [Lane 3] Embryonic (E20) rat whole brain [Lane 4] Adult rat spinal cord [Lane 5] Adult mouse whole brain [Lane 6] Adult mouse spinal cord. Rodent pNF-H protein appears as a single band of about 200 kDa in adult rat and mouse lysates, but is not present in early development (Lane 3). Additional bands appearing on the blot (Lane 4) are most likely partially degraded products of pNF-H protein.

Immunohistochemistry analysis of a formalin fixed paraffin embedded human cerebellum section with Anti-NF-H Antibody [AH1] (A85340) at a dilution of 1:4,000 detected with DAB (brown) using the Vector Labs ImmPRESS method and reagents with citra buffer retrieval. Counterstained with Hematoxylin (blue). Anti-NF-H Antibody [AH1] (A85340) strongly labels basket cell processes and certain parallel fibers and also axons in the molecular and granular layers. Note: this antibody performs well in testing with both 4% PFA and standard NBF fixed rat, mouse and human tissues.

Immunohistological analysis of rat cerebellum section stained with Anti-Neurofilament Heavy Polypeptide Antibody [AH1] (A85340), dilution 1:2,000, in green, and co-stained with Anti-Fox3 Antibody (A85403), dilution 1:5,000, in red. Following transcardial perfusion with 4% paraformaldehyde, the brain was post-fixed for 24 hours, cut to 45 µm, and free-floating sections were stained with the above antibodies. The Anti-Neurofilament Heavy Polypeptide Antibody [AH1] stains axons in the granular layer and white matter and prominent basket cell axons surrounding the large Purkinje neurons. The FOX3 antibody specifically labels nuclei of granular and other neurons, but does not stain Purkinje cells.