NF-H Antibody (AH1) - BSA Free

Western Blot: NF-H Antibody (AH1) [NBP1-05209] - Analysis of the heavily phosphorylated axonal form of NF-H protein (pNF-H) in neural tissue lysates (20ug/lane) with affinity purified mouse monoclonal anti-pNF-H antibody (NBP1-05209) at dilution of 1:5,000. Lanes on the blot are: [1] Protein size marker, [2] Adult rat whole brain [3] Embryonic (E20) rat whole brain [4] Adult rat spinal cord [5] Adult mouse whole brain [6] Adult mouse spinal cord. Rodent pNF-H protein appears as a single band of about 200kDa 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.

Immunocytochemistry/Immunofluorescence: NF-H Antibody (AH1) [NBP1-05209] - Mixed neuron/glial cultures stained with NBP1-05209 (green) and also stained with rabbit polyclonal antibody to neurofilament NF-L NB300-131 (red). The NF-L antibody stains neurofilaments in both axons and dendrites, and so can reveal neuronal cell bodies, while NBP1-05209 binds to only heavily phosphorylated forms of NF-H which are localized to mature axonal neurofilaments. In this image a few axons course from left to right and top to bottom since they contain both NF-L and phosphorylated NF-H they appear golden in color. Blue shows the distribution of DNA.

Immunohistochemistry Free-Floating: NF-H Antibody (AH1) [NBP1-05209] - Analysis of rat cerebellum section stained with mouse mAb to pNF-H, NBP1-05209, dilution 1:2,000 in green, and costained with rabbit pAb to FOX3/NeuN, dilution 1:5,000 in red. Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45uM, and free-floating sections were stained with above antibodies. The NBP1-05209 antibody stains axons in the granular layer and white matter and prominent basket cell axons surrounding the large Purkinje neurons. The FOX3/NeuN antibody specifically labels nuclei of granular and other neurons, but does not stain Purkinje cells.

• Reactivity: Hu, Mu, Rt, Po, Bv, Eq
• Applications: WB, ICC/IF, IHC, IHC
• Clone: AH1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

NF-H Antibody (AH1) - BSA Free Summary

• Description: Novus Biologicals Mouse NF-H Antibody (AH1) - BSA Free (NBP1-05209) is a monoclonal antibody validated for use in IHC, WB and ICC/IF. Anti-NF-H Antibody: Cited in 1 publication. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Native axonal phosphorylated Neurofilament Heavy protein purified from bovine spinal cord
• Marker: Neuronal Marker
• Specificity: Clone AH1 recognizes phosphorylated NF-H KSP sequences. Does not recognize non-phosphorylated KSP sequences.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: NEFH
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:1000
  • Immunohistochemistry Free-Floating 1:1000
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Paraffin
  • Western Blot 1:10000
• Application Notes: This 200kDa Neurofilament Heavy antibody is useful for Imunocytochemistry, Western Blot, and Immunohistochemistry (both IHC-P and IHC-Fr). Use in IHC-P reported in scientific literature (PMID:35835391).
• Theoretical MW: 200 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

In some species there is some cross-reactivity with the phosphorylated KSP sequences found in the related neurofilament subunit NF-M. The antibody recognizes NF-H strongly in all mammals tested to date and also in chicken

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% glycerol
• Preservative: 5mM Sodium Azide
• Concentration: 1 mg/ml
• Purity: Affinity purified

Alternate Names for NF-H Antibody (AH1) - BSA Free

• 200 kDa neurofilament protein
• KIAA0845
• NEFH
• Neurofilament Heavy (200kDa)
• neurofilament heavy polypeptide
• Neurofilament triplet H protein
• neurofilament, heavy polypeptide 200kDa
• neurofilament, heavy polypeptide
• NF200
• NFH
• NF-H

Background

Neuronal intermediate filaments (NIF) are found specifically on neurons where they play key role in maintaining neuronal morphology as well as in regenerating myelinated axons. NIF contain three subunits: a light polypeptide (NEFL/NFL), a medium polypeptide (NEFM/NFM), and a heavy polypeptide (NEFH/NFH), with molecular weights of 68, 160, and 212 kD respectively. 200kD neurofilament heavy (NEFH) is one of the major components of neuronal cytoskeleton NIF and NEFH gene was originally proposed as a DNA marker for presymptomatic diagnosis in neurofibromatosis type 2 (NF2). NEHF facilitate interactions between NIF and brain mitochondria with binding efficiency dependent upon mitochondrial membranes potential. NEFH and beta-catenin are constituents in postsynaptic density (PSD) in CNS and NEFH is essential for maintaining normal cell integrity. Diminished NEFH levels are seen in certain cancers including human autonomic nerve tumors or central neurocytomas, prostate cancer, clear-cell epithelioid tumor and small cell lung carcinoma. Fibroblasts overexpressing NEFH are distorted, multinucleated, and possess inclusions, typical changes seen in neuropathies, and defects in NEFH are linked to susceptibility to ALS.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for NF-H Antibody (NBP1-05209)(1)

Publication using NBP1-05209

• Duan S, Sawyer TW, Sontz RA et al. GFAP-directed Inactivation of Men1 Exploits Glial Cell Plasticity in Favor of Neuroendocrine Reprogramming Cellular and molecular gastroenterology and hepatology 2022-07-11 [PMID: 35835391] (IHC-P, Mouse)

Bioinformatics

• Gene Symbol: NEFH
• Entrez:
  • 528842(Bovine)
  • 4744(Human)
  • 380684(Mouse)
  • 24587(Rat)