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NF-H Antibody

Catalog # NB300-135 | Novus Biologicals a Bio-Techne Brand
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NB300-135

Key Product Details

Species Reactivity

Human, Mouse, Rat, Porcine, Bovine, Equine

Applications

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, In vitro assay, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Product Summary for NF-H Antibody

Immunogen

Purified bovine Neurofilament Heavy protein.

Specificity

Phosphorylated and non-phosphorylated forms of 200kDa Neurofilament Heavy

Marker

Neuronal Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

210 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

Western Blot: NF-H Antibody [NB300-135] - Proof of viability of organotypic adult brain slices. Brain slices of adult 9 month old wildtype (WT) mice were dissected fresh or incubated for either 1 day or for 14 days to examine vitality of slices (A). Brain slices were subsequently analyzed by Western blot using antibodies against Neurofilament-200 kDa, total tau (Tau-5) and actin served as a control. Size markers are given as kDa on the right side. Note some degradation of neurofilament but stability of Tau-5. Image collected and cropped by CiteAb from the following publication (http://journal.frontiersin.org/article/10.3389/fnagi.2018.00113/full) licensed under a CC-BY license.
Immunohistochemistry: NF-H Antibody [NB300-135] - Mouse hippocampus section stained with rabbit pAb to NF-H, dilution 1:2,000 in red, and costained with mouse mAb to myelin basic protein (MBP), dilution 1:5,000 in green. The blue is DAPI staining of nuclear DNA. 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 NF-H antibody labels a network of axons of different neurons, while the MBP antibody stains myelin sheath around these axons.
Immunohistochemistry: NF-H Antibody [NB300-135] - Immunostainings of adult organotypic brain slices at the hippocampal level. Brain slices were prepared from WT or TG adult mice and incubated with okadaic acid (OA). Brain slices were stained for nuclear DAPI (blue, A,B) showing intact brain structures. Thiazine Red staining (C,D) shows red fluorescent plaques in the hippocampus (D) of TG mice, while only background is seen in the hippocampus (C) of WT mice. Staining of neurofilament fibers (G,H) shows good neuronal viability both in WT and TG mice. Image collected and cropped by CiteAb from the following publication (http://journal.frontiersin.org/article/10.3389/fnagi.2018.00113/full) licensed under a CC-BY license.

Applications for NF-H Antibody

Application
Recommended Usage

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

1:1000-1:5000

Immunohistochemistry

1:1000-1:5000

Immunohistochemistry-Frozen

1:1000-1:5000

Immunohistochemistry-Paraffin

1:1000-1:5000

Western Blot

1:10000-1:25000
Application Notes
This 200kDa Neurofilament Heavy Antibody is useful for WB, ICC/IF, ELISA and IHC-P, IHC-Frozen. In WB a band can be seen at around 200kDa corresponding to rodent Neurofilament Heavy. In larger species such as porcine or human, Neurofilament Heavy runs a little slower and can be seen around 210-220kDa. 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. Use in In vitro assay reported in scientific literature (PMID 27789413). Use in IHC-WhMt reported in scientific literature (PMID:33499508)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Published Applications

Read 28 publications using NB300-135 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Supplied as serum

Preservative

5mM Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

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

Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called Neurofilament Light (NF-L), Neurofilament Medium (NF-M) and Neurofilament Heavy (NF-H). Neurofilament Heavy is the heavy or high molecular weight microfilament subunit and runs on SDS-PAGE gels in the range 180-220kDa, with some variation in different species. Neurofilament Heavy antibodies can be used to identify neurons and their processes in tissue sections and in tissue culture. Additionally, antibodies can also be used to study microfilament accumulations seen in many neurological diseases, such as Lou Geri's disease or Alzheimer's disease.

Long Name

Neurofilament Protein, Heavy Chain

Alternate Names

NEFH, NFH

Entrez Gene IDs

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

Gene Symbol

NEFH

Product Documents for NF-H Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for NF-H Antibody

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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FAQs for NF-H Antibody

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  • Q: I am using the 200kDa Neurofilament Heavy for the first time. Would it be appropriate to mix it with glycerol for freezer storage rather than aliquot, and do you recommend any antigen retrieval?

    A: This product is unpurified whole antisera and should be stable at 4C for few weeks to 6 months, because it contains Sodium Azide as antimicrobial agent (prevents bacterial contamination of the serum). However, if you plan to store it for longer period, you can divide it into aliquots and store the same at -20C or -80C. We do not have a customized protocol for the IHC-P application for this product, however, in general the antigen retrieval step is optional. If you are working on archival tissues or have fixed the tissues for prolonged period, I would highly suggest you to include the antigen retrieval step to your protocol. Louis et al 2012 have cited the use of this antibody for IHC-P application and they have included antigen retrieval step in their experiments. You may follow this paper for more details and images for compression with your study: Louis ED, et al. Neurofilament protein levels: quantitative analysis in essential tremor cerebellar cortex. Neurosci Lett. 2012 Jun 14;518(1):49-54. Epub 2012 May 4. PMID: 22561033

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