NF-H Antibody (AH1)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-05209

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Equine

Cited:

Mouse

Applications

Validated:

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Free-Floating, Immunohistochemistry-Paraffin, Western Blot

Cited:

Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # AH1

Concentration

1 mg/ml

View all NF-H Antibodies »

Product Specifications

Immunogen

Native axonal phosphorylated Neurofilament Heavy protein purified from bovine spinal cord

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

Specificity

Clone AH1 recognizes phosphorylated NF-H KSP sequences. Does not recognize non-phosphorylated KSP sequences.

Marker

Neuronal Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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.

Scientific Data Images for NF-H Antibody (AH1)

Western Blot: NF-H Antibody (AH1) [NBP1-05209]

Western Blot: NF-H Antibody (AH1) [NBP1-05209]

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]

Immunocytochemistry/ Immunofluorescence: NF-H Antibody (AH1) [NBP1-05209]

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]

Immunohistochemistry Free-Floating: NF-H Antibody (AH1) [NBP1-05209]

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.

Applications for NF-H Antibody (AH1)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:1000

Immunohistochemistry

1:1000

Immunohistochemistry Free-Floating

1:1000

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).

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

50% PBS, 50% glycerol

Preservative

0.035% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: NF-H

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.

Long Name

Neurofilament Protein, Heavy Chain

Alternate Names

NEFH, NFH

Entrez Gene IDs

4744 (Human); 380684 (Mouse); 24587 (Rat); 528842 (Bovine)

Gene Symbol

NEFH

Additional NF-H Products

Product Documents for NF-H Antibody (AH1)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for NF-H Antibody (AH1)

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.

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