NF-M Antibody

Western Blot: NF-M Antibody [NB300-222] - Western blot analysis of different neuronal tissue and cell lysates using chicken pAb to NF-M, NB300-222, dilution 1:2,000 in green: [1] protein standard (red), [2] rat brain [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, [6] NIH/3T3 cells, [7] HEK293, [8] HeLa, [9] SH-SY5Y, and [10] C6 cells. Strong band at 145kDa corresponds to rodent NF-M, and about 160kDa band corresponds to human NF-M protein, visible in SHSY-5Y and HEK293 cells which have neuronal properties. NF-M is not expressed in HeLa and other cell lines tested.

Immunocytochemistry/Immunofluorescence: NF-M Antibody [NB300-222] - 160kDa Neurofilament Medium Antibody [NB300-222] - View of mixed neuron/glial cultures stained with NB300-222 (red). The NF-M protein is assembled into neurofilaments which are found throughout the axons, dendrites and perikarya of these cells.

Immunohistochemistry Free-Floating: NF-M Antibody [NB300-222] - Immunofluorescent analysis of rat cerebellum section stained with chicken pAb to NF-M, NB300-222, dilution 1:1,000 in red, and costained with mouse mAb to CNPase, dilution 1:500, in green. The blue is DAPI staining of nuclear DNA. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45uM, and free floating sections were stained with above antibodies. The NF-M antibody labels the axons of basket cells and other neurons, while the CNP antibody stains oligodendrocytes, cells that form the myelin sheathes around axons.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ch, Dr, Eq
• Applications: WB, ICC/IF, IHC, WB, IHC, KD
• Clonality: Polyclonal
• Host: Chicken
• Conjugate: Unconjugated

NF-M Antibody Summary

• Immunogen: Recombinant construct containing the C-terminus of the human sequence (amino acids 708-877) expressed in and purified from E. coli.
• Marker: Neuronal Marker
• Isotype: IgY
• Clonality: Polyclonal
• Host: Chicken
• Gene: NEFM
• Purity: IgY purified

Applications/Dilutions

• Dilutions:
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:1000
  • Immunohistochemistry Free-Floating
  • Immunohistochemistry 1:1000
  • Knockdown Validated
  • Western Blot 1:10000
• Application Notes: This 160kDa Neurofilament Medium antibody is useful for ICC/IF, IHC and Western blot. In Western blot this antibody detects a band at ~145-160kDa. Knockdown validation (PMID: 31811140). Use in immunoblotting reported in scientific literature (PMID:31811140).
• Theoretical MW: 160 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.
• Control:

  NF-M Overexpression Lysate

Reactivity Notes

Drosophila reactivity reported in scientific literature (PMID:31811140).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Supplied as an aliquot of IgY preparation
• Preservative: 5mM Sodium Azide
• Purity: IgY purified

Notes

Chicken products cannot be exported to Canada.

Alternate Names for NF-M Antibody

• NEF3
• NEF3neurofilament medium polypeptide
• NEFM
• Neurofilament 3
• neurofilament, medium polypeptide 150kDa
• neurofilament, medium polypeptide
• neurofilament-3 (150 kD medium)
• NFM
• NF-M
• NF-M160 kDa neurofilament protein
• NFMNeurofilament triplet M protein

Background

Neuronal intermediate filaments (NIF) are type-IV intermediate filaments 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. Mature NIFs are heteropolymers with about 4:2:1 stoichiometric ratio of NEFL: NEFM: NEFH and also contain a fourth type-IV intermediate filament (IF) subunit, alpha-internexin. NEFM is essential to cross-bridge formation, stabilization, and longitudinal extension of filamentous network and is more critical than NEFH in regulating NIF's structure and function. NEFM is crucial for the acquisition of normal axonal caliber in response to a myelin-dependent "outside-in" trigger for radial axonal growth and removal of the tail domain as well as lysine-serine-proline (KSP) repeats of NEFM, but not NEFH, produces axons with impaired radial growth and reduced conduction velocities. Moreover, mutations of NEFM have been associated with CNS/PNS disorders such as ALS, Parkinson's disease etc., and an imbalance in NEFM's O-GlcNAcylation - phosphorylation regulation plays a crucial role in in Alzheimer 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-M Antibody (NB300-222)(10)

Publications using NB300-222

• Rutigliani C, Tribble JR, HagstrOm A et al. Widespread retina and optic nerve neuroinflammation in enucleated eyes from glaucoma patients Acta neuropathologica communications 2022-08-19 [PMID: 35986368] (IHC-P, Human)
  
  Details:
  Dilution used 1:400, optic nerve head sections
• Batool S, Akhter B, Zaidi J Et al. Neuronal Menin Overexpression Rescues Learning and Memory Phenotype in CA1-Specific alpha 7 nAChRs KD Mice Cells 2021-11-24 [PMID: 34943798] (ICC/IF, Mouse)
• Batool S, Zaidi J, Akhter B et al. Spatiotemporal Patterns of Menin Localization in Developing Murine Brain: Co-Expression with the Elements of Cholinergic Synaptic Machinery Cells 2021-05-16 [PMID: 34065662] (ICC/IF, Mouse)
• Iqbal F, Pehar M, Thompson AJ et al. A synthetic peptide rescues rat cortical neurons from anesthetic-induced cell death, perturbation of growth and synaptic assembly Scientific reports 2021-02-25 [PMID: 33633281] (ICC/IF, Rat)
• Casci I, Krishnamurthy K, Kour S Muscleblind acts as a modifier of FUS toxicity by modulating stress granule dynamics and SMN localization Nat Commun 2019-12-06 [PMID: 31811140] (IB, ICC/IF, KD, Human, Drosophila, Rat)
• Chu TH, Cummins K, Sparling JS et al. Axonal and myelinic pathology in 5xFAD Alzheimer's mouse spinal cord. PLoS ONE 2017-11-27 [PMID: 29176903] (Mouse)
• Getz AM, Xu F, Visser F et al. Tumor suppressor menin is required for subunit-specific nAChR a5 transcription and nAChR-dependent presynaptic facilitation in cultured mouse hippocampal neurons. Sci Rep 2017-05-11 [PMID: 28496137] (ICC/IF, Mouse)
• Xu F, Armstrong R, Urrego D et al. The mitochondrial division inhibitor Mdivi-1 rescues mammalian neurons from anesthetic-induced cytotoxicity Mol Brain 2016-03-24 [PMID: 27009068] (ICC/IF, Rat)
• Gold M, Mengel D, Roskam S, Dodel R, Bach JP. Mechanisms of action of naturally occurring antibodies against beta-amyloid on microglia. J Neuroinflammation 2013-01-14 [PMID: 23317003] (WB, Mouse)
• Portmann-Lanz CB, Schoeberlein A, Portmann R et al. Turning placenta into brain: placental mesenchymal stem cells differentiate into neurons and oligodendrocytes. Am J Obstet Gynecol. 2010-03-01 [PMID: 20060088] (ICC/IF, Human)

Review for NF-M Antibody (NB300-222) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Verified Customer ICC Rat 10/17/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Cortical neurons
• Species: Rat
• Lot: 1-0104

Comments

• Comments: Confocal microscope image of cultured rat cortical (14 days invitro) neurons fixed with 4% paraformaldehyde and stained for Neurofilament (red) and MAP-2 (green). Neurofilament antibody was used at a dilution of 1:1000 and incubated at 4 degrees overnight.

Bioinformatics

• Gene Symbol: NEFM
• Entrez:
  • 4741(Human)
  • 24588(Rat)
• Uniprot:
  • P07197()