alpha-Internexin Antibody

Immunohistochemistry: alpha-Internexin Antibody [NB300-139] - Rat cerebellum section stained with rabbit pAb to alpha-internexin, dilution 1:2,000, in green, and chicken pAb to GFAP, dilution 1:5,000, in red. 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 alpha-internexin antibody selectively stains axons and dendrites of neuronal cells, in particular Purkinje cells and parallel fibers the axons of granule cells. The GFAP antibody labels network of glial cells, such as astrocytes in the granule cell layer and white matter and Bergmann glia in the molecular layer.

Western Blot: alpha-Internexin Antibody [NB300-139] - Whole tissue lysates using rabbit pAb to alpha-internexin, dilution 1:10,000 in green: [1] protein standard (red), [2] mouse spinal cord, [3] rat spinal cord, [4] bovine spinal cord. Major bands in the 64-66 kDa range corresponds to alpha-internexin. The alpha-internexin protein from different species is known to vary slightly in SDS-PAGE molecular weight.

Western Blot: alpha-Internexin Antibody [NB300-139] - The expression of a-internexin (INA) was analyzed by Western blotting. White bars are controls, grey bars are enucleated and black bars are anophthalmic mice. Results from V1 (left), V2M (middle) and V2L (right) are shown. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0159320) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: alpha Internexin Antibody [NB300-139] - IF Confocal analysis of SHSY5Y cells using alpha Internexin antibody (NB300-139, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).

Western Blot: alpha Internexin Antibody [NB300-139] - Western blot of whole rat spinal cord homogenate stained with RPCA-a-Int, at dilution of 1:20,000. A prominent band running at ~66kDa is apparent, as well as smaller lower bands which are apparently degradation products. A minor band at ~150kDa is also seen, apparently resulting from dimerization of alpha-internexin.

Immunocytochemistry/Immunofluorescence: alpha Internexin Antibody [NB300-139] - Mixed neuron-glial cultures stained with rabbit antibody to alpha-internexin (red) and chicken antibody to peripherin CPCA-Peri (green). The alpha internexin antibody stains numerous axonal and dendritic profiles in these cultures, while peripherin antibody binds to only a subset of neurons.

Immunocytochemistry/Immunofluorescence: alpha Internexin Antibody [NB300-139] - Polyclonal alpha internexin, NB 300-139 (red) staining neuronal progenitor cells. The green stain shows the fibroblast marker, Plectin (not one of our antibodies).

Proteins involved in morphogenesis.The expression of a-internexin (INA), collapsing response mediator protein 2 (CRMP2) and 4 (CRMP4), neurofilament-low (NFL) and tubuline-b2 (Tub beta 2) were analyzed by Western blotting. White bars are controls, grey bars are enucleated and black bars are anophthalmic mice. Results from V1 (left), V2M (middle) and V2L (right) are shown for each protein. * P < 0.05 and ** P < 0.01.

Western Blot: alpha-Internexin Antibody [NB300-139] - Proteins involved in morphogenesis.The expression of a-internexin (INA), collapsing response mediator protein 2 (CRMP2) & 4 (CRMP4), neurofilament-low (NFL) & tubuline-b2 (Tub beta 2) were analyzed by Western blotting. White bars are controls, grey bars are enucleated & black bars are anophthalmic mice. Results from V1 (left), V2M (middle) & V2L (right) are shown for each protein. * P < 0.05 & ** P < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27410964), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Eq
• Applications: WB, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

alpha-Internexin Antibody Summary

• Description: Novus Biologicals Rabbit alpha-Internexin Antibody (NB300-139) is a polyclonal antibody validated for use in IHC, WB, ICC/IF and IP. Anti-alpha-Internexin Antibody: Cited in 10 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Purified recombinant rat alpha internexin expressed in and purified from E. coli. [UniProt# P23565]
• Localization: Cytoplasmic
• Marker: Immature Neuronal Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: INA
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:500 - 1:1000
  • Immunohistochemistry 1:500 - 1:1000
  • Immunoprecipitation
  • Western Blot 1:10000 - 1:20000
• Application Notes: This alpha Internexin antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western blot, where a band is observed ~ 66kDa. Use in IP was reported in the scientific literature (PMID: 23802559).
• Theoretical MW: 66 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: 0.035% Sodium Azide
• Purity: Unpurified

Alternate Names for alpha-Internexin Antibody

• 66 kDa neurofilament protein
• alphaInternexin
• alpha-Internexin
• alpha-Inx
• FLJ18662
• FLJ57501
• INA
• internexin neuronal intermediate filament protein, alpha
• MGC12702
• NEF5
• neurofilament 5 (66kD)
• Neurofilament 5
• Neurofilament-66
• neurofilament-66, tax-binding protein
• NF-66
• NF-66alpha-internexin
• TXBP-1

Background

In mammalian CNS, five key neuronal intermediate filament (NIF) proteins have been discovered which includes alpha-internexin (66 kD, INA or Inexa), peripherin (57 kD), and three neurofilament (NF) proteins, which are neurofilament light (NF-L, 68 kD), medium (NF-M, 145 kD), and heavy (NF-H, 200 kD). Among NIFs, alpha-internexin is categorized as class IV NIF which is highly expressed in most neurons during development especially when they begin to differentiate and before the expression of NF triplet proteins, and it is structurally/functionally associated with NF triplet proteins inside mature CNS neurons. Peripherin is mainly expressed in PNS and in some CNS neurons also, and alpha-internexin - peripherin self-assemble or co-assemble with neurofilament protein subunits to form filamentous structure before their translocation into the axons to become an essential constituent of shape-maintaining IF network in mature neurons. It has been recognized as one of the many proteins phosphorylated by ATM/ATR in response to DNA damage and has been associated with developmental, degenerative /overexpression causes neuronal cell death, and inflammatory processes of CNS. Abnormal NIF accumulation is a neuropathological signature of several neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, and amyotrophic lateral sclerosis. Alpha-internexin has been detected as a major component of pathological inclusions in frontotemporal dementia, also called 'neuronal intermediate filament inclusion disease (NIFID).

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 alpha-Internexin Antibody (NB300-139)(10)

Publications using NB300-139

• Huynh DT, Hu J, Schneider JR et al. O-GlcNAcylation regulates neurofilament-light assembly and function and is perturbed by Charcot-Marie-Tooth disease mutations bioRxiv : the preprint server for biology 2023-02-22 [PMID: 36865196]
• Atkinson R, Leung J, Bender J, et al. TDP-43 mislocalization drives neurofilament changes in a novel model of TDP-43 proteinopathy Disease models & mechanisms 2021-01-06 [PMID: 33408125] (IF/IHC, Mouse)
• Laramee ME, Smolders K, Hu TT et al. Congenital Anophthalmia and Binocular Neonatal Enucleation Differently Affect the Proteome of Primary and Secondary Visual Cortices in Mice. PLoS One 2016-07-13 [PMID: 27410964] (WB)
• Blizzard CA, King AE, Vickers J, Dickson T. Cortical murine neurons lacking the neurofilament light chain protein have an attenuated response to injury in vitro. J Neurotrauma 2013-06-26 [PMID: 23802559] (IP, Mouse)
• Liu Y, Staal JA, Canty AJ et al. Cytoskeletal changes during development and aging in the cortex of neurofilament light protein knockout mice. J Comp Neurol 2013-06-01 [PMID: 23172043] (IF/IHC, ICC/IF, WB, Mouse)
• King AE, Blizzard CA, Southam KA, Vickers JC, Dickson TC. Degeneration of axons in spinal white matter in G93A mSOD1 mouse characterized by NFL and alpha-internexin immunoreactivity. Brain Res. 2012-05-17 [PMID: 22609817] (IF/IHC, ICC/IF, Mouse)
• Blizzard CA, Chuckowree JA, King AE et al. Focal Damage to the Adult Rat Neocortex Induces Wound Healing Accompanied by Axonal Sprouting and Dendritic Structural Plasticity. Cereb Cortex. 2010-05-28 [PMID: 20511339] (IF/IHC, Rat)
• King AE, Dickson TC, Blizzard CA et al. Neuron-glia interactions underlie ALS-like axonal cytoskeletal pathology Neurobiol Aging 2011-03-01 [PMID: 19427060] (IF/IHC, ICC/IF, Mouse)
• King AE, Dickson TC, Blizzard CA et al. Excitotoxicity mediated by non-NMDA receptors causes distal axonopathy in long-term cultured spinal motor neurons. Eur J Neurosci. 2007-10-01 [PMID: 17908171] (WB, ICC/IF, Rat)
• Ganns D, Schrodl F, Neuhuber W, Brehmer A. Investigation of general and cytoskeletal markers to estimate numbers and proportions of neurons in the human intestine. Histol Histopathol 2006-01-01 [PMID: 16267786] (Human)

Review for alpha-Internexin Antibody (NB300-139) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Bryan Tinsley ICC Human 06/16/2014

Summary

• Application: Immunocytochemistry
• Species: Human

Bioinformatics

• Gene Symbol: INA
• Entrez:
  • 9118(Human)
  • 24503(Rat)
• Uniprot:
  • P23565(Rat)