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 (//journal.frontiersin.org/article/10.3389/fnagi.2018.00113/full) licensed under a CC-BY licence.

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 (//journal.frontiersin.org/article/10.3389/fnagi.2018.00113/full) licensed under a CC-BY licence.

Western Blot: NF-H Antibody [NB300-135] - Analysis of 200kDa Neurofilament Heavy expression in rat spinal cord extract. The first lane is Coomassie Brilliant Blue stained and the second lane is probed with rabbit anti-Neurofilament Heavy antibody NB300-135. The NF-H corresponds to a weight of 200kDa.

Western Blot: NF-H Antibody [NB300-135] - Different tissue lysates using rabbit pAb to NF-H, dilution 1:10,000 in green: [1] protein standard (red), [2] rat brain, [3] rat spinal cord [4] mouse brain, and [5] mouse spinal cord lysate. Strong band at about 220 kDa corresponds to the phosphorylated axonal form of the NF-H subunit. Smaller proteolytic fragments of NF-H are also detected with RPCA-NF-H antibody.

• Reactivity: Hu, Mu, Rt, Po, Bv, Eq
• Applications: WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P, In vitro, IHC-WhMt
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

NF-H Antibody Summary

• Immunogen: Purified bovine Neurofilament Heavy protein.
• Marker: Neuronal Marker
• Specificity: Phosphorylated and non-phosphorylated forms of 200kDa Neurofilament Heavy
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NEFH
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100-1:2000
  • Immunocytochemistry/Immunofluorescence 1:1000-1:5000
  • Immunohistochemistry Whole-Mount
  • Immunohistochemistry 1:1000-1:5000
  • Immunohistochemistry-Frozen 1:1000-1:5000
  • Immunohistochemistry-Paraffin 1:1000-1:5000
  • In vitro assay
  • 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)
• 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Supplied as serum
• Preservative: 5mM Sodium Azide
• Purity: Unpurified

Alternate Names for NF-H Antibody

• 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

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.

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 (NB300-135)(22)

Publications using NB300-135

• Manto KM, Govindappa PK, Martinazzi B et al. Erythropoietin-PLGA-PEG as a local treatment to promote functional recovery and neurovascular regeneration after peripheral nerve injury Journal of nanobiotechnology 2022-10-28 [PMID: 36307805] (WB)
• Ucar B, Stefanova N, Humpel C Spreading of Aggregated alpha-Synuclein in Sagittal Organotypic Mouse Brain Slices Biomolecules 2022-01-19 [PMID: 35204664] (WB, Mouse)
• Liu W, Jiang H, Liu X Et Al. Melatonin Abates TMJOA Chronic Pain by MT2R in Trigeminal Ganglion Neurons Journal of dental research 2021-07-27 [PMID: 34315312]
• Manto KM, Govindappa PK, Parisi D Et al. (4-Aminopyridine)PLGAPEG as a Novel Thermosensitive and Locally Injectable Treatment for Acute Peripheral Nerve Injury ACS Appl Bio Mater 2021-06-18 [PMID: 34142019]
• Ulger M, Sezer G, Ozyazgan I et al. The effect of erythropoietin and umbilical cord-derived mesenchymal stem cells on nerve regeneration in rats with sciatic nerve injury Journal of chemical neuroanatomy 2021-04-14 [PMID: 33864937] (IF/IHC, Rat)
• Moelgg K, Jummun F, Humpel C Spreading of Beta-Amyloid in Organotypic Mouse Brain Slices and Microglial Elimination and Effects on Cholinergic Neurons Biomolecules 2021-03-15 [PMID: 33804246] (WB, Mouse)
• Lee JI, Gurjar AA, Talukder MAH, et al. Purposeful Misalignment of Severed Nerve Stumps in a Standardized Transection Model Reveals Persistent Functional Deficit With Aberrant Neurofilament Distribution Military medicine 2021-01-25 [PMID: 33499508] (IHC-WhMt, Mouse)
• Jiang H, Xu L, Liu W et al. The peripheral and central expression of CGRP and IB4 in chronic pain from MIA-induced TMJOA rats Cell Mol Neurobiol 2021-01-02 [PMID: 33387118]
• Aharoni R, Globerman R, Eilam R et al. Titration of myelin oligodendrocyte glycoprotein (MOG) - Induced experimental autoimmune encephalomyelitis (EAE) model Cells 2020-11-09 [PMID: 33189793] (WB, Human)
• Govindappa Pk, Talukder Mah, Gurjar Aa et Al. An effective erythropoietin dose regimen protects against severe nerve injury-induced pathophysiological changes with improved neural gene expression and enhances functional recovery Int. Immunopharmacol. 2020-03-03 [PMID: 32143001] (IHC-P, IF/IHC, Mouse)
• Tomlinson J, Zwirner J, Ondruschka B et al. Innervation of the hip joint capsular complex: A systematic review of histological and immunohistochemical studies and their clinical implications for contemporary treatment strategies in total hip arthroplasty PLoS ONE 2020-02-26 [PMID: 32101545] (IF/IHC)
• Long-term culturing of porcine nodose ganglia Kuan SP, Atanasova KR, Guevara MV J Neurosci Methods [PMID: 31821820] (ICC/IF, Porcine)
• Hutter-Schmid B, Humpel C. Primary mouse brain pericytes isolated from transgenic Alzheimer mice spontaneously differentiate into a CD11b+ microglial-like cell type in vitro Exp. Gerontol. 2018-08-09 [PMID: 30099090] (WB, Mouse)
• Foidl BM, Humpel C. Differential Hyperphosphorylation of Tau-S199, -T231 and -S396 in Organotypic Brain Slices of Alzheimer Mice. A Model to Study Early Tau Hyperphosphorylation Using Okadaic Acid. Front Aging Neurosci 2018-04-19 [PMID: 29725295] (Mouse)
• Ermilov AN, Kumari A, Li L et al. Maintenance of Taste Organs Is Strictly Dependent on Epithelial Hedgehog/GLI Signaling. PLoS Genet. 2016-11-01 [PMID: 27893742] (WB, Mouse)
• Skrzypczyk A, Giri S, Bader A. Generation of induced pluripotent stem cell line from foreskin fibroblasts Stem Cell Research 2016-09-15 [PMID: 27789413] (In vitro, Human)
• Kumari A, Ermilov An, Allen Bl et al. Hedgehog pathway blockade with the cancer drug LDE225 disrupts taste organs and taste sensation. J. neurophysiol. 2014-11-12 [PMID: 25392175]
• From R, Eilam R, Bar-Lev DD et al. Oligodendrogenesis and myelinogenesis during postnatal development effect of glatiramer acetate. Glia 2014-02-13 [PMID: 24481644] (IHC-P, Mouse)
• Liu HX, Ermilov A, Grachtchouk M et al. Multiple Shh signaling centers participate in fungiform papilla and taste bud formation and maintenance. Dev Biol 2013-08-02 [PMID: 23916850] (IF/IHC, Mouse)
• Elan D Louis, Karen, Rachel Babij, Etty Cortes, Ronald K Liem, Jean-Paul G Vonsattel, Phyllis L Faust. Neurofilament Protein Levels: Quantitative Analysis in Essential Tremor Cerebellar Cortex, . Neuroscience Letters, Available online , 10.1016/j.neulet.2012.04.054. 2012-05-04 [PMID: 22561033] (IF/IHC, WB, Human)
• O'Donnell LA, Agrawal A, Sabnekar P et al. Apelin, an endogenous neuronal peptide, protects hippocampal neurons against excitotoxic injury J Neurochem. 2007-09-01 [PMID: 17767704] (ICC/IF, Rat)
• O Donnell, LA et al. Human immunodeficiency virus (HIV)-induced neurotoxicity roles for the NMDA receptor subtypes. J Neurosci; 26:981-990. 2006-01-01 [PMID: 16421318] (WB, Human)

FAQs for NF-H Antibody (NB300-135). (Showing 1 - 1 of 1 FAQs).

1. 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?
   • 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

Bioinformatics

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