alpha-Internexin Antibody (2E3) - BSA Free

Western Blot: alpha-Internexin Antibody (2E3) [NB300-140] - Analysis of different tissue lysates using alpha-Internexin, dilution 1:10000 (Red): [1] protein standard, [2] rat brain, [3] rat spinal cord, [4] mouse brain, [5] mouse spinal cord, and [6] cow spinal cord lysate. The antibody reveals the alpha-Internexin protein with an apparent molecular weight of 64-66kDa, with some variability across species.

Immunocytochemistry/Immunofluorescence: alpha-Internexin Antibody (2E3) [NB300-140] - Hippocampal neurons in tissue culture with neurofilament alpha internexin (red), using NB300-140 and GFAP (green), using NB300-141.

Immunohistochemistry: alpha-Internexin Antibody (2E3) [NB300-140] - Section of rat facial nucleus 7 days following axotomy. These neurons are capable of regenerating their axons and also, concomitant with regeneration, strongly upregulate alpha-Internexin in their perikarya. Other central neurons which are not able to regenerate their axons do not upregulate this protein after axotomy and untreated facial neurons normally show only very low levels of alpha-Internexin. Both findings suggest that alpha-Internexin has a role in axonal regeneration.

Simple Western: alpha-Internexin Antibody (2E3) [NB300-140] - Simple Western lane view shows a specific band for alpha Internexin in 0.5 mg/ml of SH-SY5Y lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po, Bv
• Applications: WB, Simple Western, ICC/IF, IHC
• Clone: 2E3
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

alpha-Internexin Antibody (2E3) - BSA Free Summary

• Immunogen: Purified recombinant rat alpha Internexin expressed in and purified from E. coli [UniProt# P23565]
• Epitope: The C-terminal non-helical extension of the protein.
• Localization: Cytoplasmic
• Marker: Immature Neuronal Marker
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: INA
• Purity: Affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:5000
  • Immunohistochemistry 1:5000
  • Immunohistochemistry-Paraffin 1:5000
  • Simple Western 1:200
  • Western Blot 1:10000
• Application Notes: This alpha Internexin antibody is useful for Immunohistochemistry paraffin embedded sections, Immunocytochemistry/Immunofluorescence and Western blot. This antibody can be used on formalin-fixed cells in tissue culture and cryostat sections. A 66 kDa band can be seen in Western blotting. The epitope recognized by the 2E3 clone is in the C-terminal non-helical extension of the protein and is unusually resistant to aldehyde fixation, so that this antibody is ideal for studies of formalin-fixed paraffin-embedded histological sections.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in SH-SY5Y lysate, separated by Size, antibody dilution of 1:200, apparent MW was 140 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
  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.
• 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.

Reactivity Notes

Porcine reported in scientific literature (PMID: 12197769).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% Glycerol
• Preservative: 0.035% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Affinity purified

Alternate Names for alpha-Internexin Antibody (2E3) - BSA Free

• 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 (NB300-140)(34)

Publications using NB300-140

• Johnson AK, McCurdy VJ, Gray-Edwards HL et al. Life-Limiting Peripheral Organ Dysfunction in Feline Sandhoff Disease Emerges after Effective CNS Gene Therapy Annals of neurology 2023-08-01 [PMID: 37526361]
• Forest F, Masliah-Planchon J, Berger C et al. High-grade childhood intra-parenchymal brain tumor clustering with ATRT and expanding the cancer spectrum related to inherited SMARCE1 truncating variations Acta neuropathologica communications 2022-02-14 [PMID: 35164878] (IF/IHC, Human)
• Bielle F, Di Stefano AL, Meyronnet D et al. Diffuse gliomas with FGFR3-TACC3 fusion have characteristic histopathological and molecular features. Brain Pathol. 2017-10-04 [PMID: 28976058] (Human)
• Breton Q, Plouhinec H, Prunier-Mirebeau D et al. BRAF-V600E immunohistochemistry in a large series of glial and glial-neuronal tumors. Brain Behav. 2017-03-01 [PMID: 28293477] (IF/IHC, Human)
• Bielle F, Ducray F, Mokhtari K et al. Tumor cells with neuronal intermediate progenitor features define a subgroup of 1p/19q co-deleted anaplastic gliomas. Brain Pathol 2016-08-20 [PMID: 27543943]
• Matsumura N, Nobusawa S, Ikota H et al. Coexpression of cyclin D1 and alpha-internexin in oligodendroglial tumors. Brain Tumor Pathol. 2015-08-02 [PMID: 26233522] (IHC-P, Human)
• Nagaishi M, Yokoo H, Nobusawa S et al. Localized overexpression of alpha-internexin within nodules in multinodular and vacuolating neuronal tumors Neuropathology. 2015-06-14 [PMID: 26073706] (IHC-P, Human)
• Figarella-Branger D, Mokhtari K, Dehais C et al. Mitotic index, microvascular proliferation, and necrosis define 3 groups of 1p/19q codeleted anaplastic oligodendrogliomas associated with different genomic alterations. Neuro-oncology 2014-04-09 [PMID: 24723566] (IHC-P, Human)
• Reyes-Botero G, Giry M, Mokhtari K et al. Molecular analysis of diffuse intrinsic brainstem gliomas in adults. J Neurooncol. 2013-11-17 [PMID: 24242757] (IHC-P, Human)
• Nagaishi M, Suzuki A, Nobusawa S et al. Alpha-internexin and altered CIC expression as a supportive diagnostic marker for oligodendroglial tumors with the 1p/19q co-deletion. Brain Tumor Pathol. 2013-11-07 [PMID: 24197863] (IHC-P, Human)
• Takano S, Kato Y, Yamamoto T et al. Immunohistochemical detection of IDH1 mutation, p53, and internexin as prognostic factors of glial tumors J Neurooncol 2012-03-07 [PMID: 22396072] (IF/IHC, Human)
• Seilhean D, Le Ber I, Sarazin M, Lacomblez L, Millecamps S, Salachas F, Pradat PF, Le Forestier N, LeGuern E, Dubois B, Meininger V, Brice A, Hauw JJ, Duyckaerts C. Fronto-temporal lobar degeneration: neuropathology in 60 cases. J Neural Transm;118(5):753-64. 2011-05-01 [PMID: 21541771] (IF/IHC, Human)
• Mokhtari K, Ducray F, Kros J et al. Alpha-internexin expression predicts outcome in anaplastic oligodendroglial tumors and may positively impact the efficacy of chemotherapy: European organization for research and treatment of cancer trial 26951 Cancer 2011-07-01 [PMID: 21246521] (IHC-P, Human)
• San Millan B, Kaci R, Polivka M et al. Gliomatosis cerebri: A biopsy and autopsy case report Ann Pathol. 2010-02-01 [PMID: 20223351] (IF/IHC, Human)
• Wang P, Wang SM, Hsieh CJ, Chien CL. Neural expression of alpha-internexin promoter in vitro and in vivo. J Cell Biochem;97(2):275-87. 2006-02-01 [PMID: 16173078] (IF/IHC, Mouse)
• Ko TL, Chien CL, Lu KS. The expression of alpha-internexin and peripherin in the developing mouse pineal gland. J Biomed Sci;12(5):777-89. 2005-10-01 [PMID: 16132113]
• Rauch U, Klotz M, Maas-Omlor S, Wink E, Hansgen A, Hagl C, Holland-Cunz S, Schafer KH. Expression of intermediate filament proteins and neuronal markers in the human fetal gut. J Histochem Cytochem;54(1):39-46. 2006-01-01 [PMID: 16087706] (IF/IHC, ICC/IF, Human)
• Uchikado H, Shaw G, Wang DS, Dickson DW. Screening for neurofilament inclusion disease using alpha-internexin immunohistochemistry. Neurology;64(9):1658-9. 2005-05-10 [PMID: 15883343] (IF/IHC, Human)
• Chien CL, Liu TC, Ho CL, Lu KS. Overexpression of neuronal intermediate filament protein alpha-internexin in PC12 cells. J Neurosci Res;80(5):693-706. 2005-06-01 [PMID: 15880430] (WB, ICC/IF, Rat)
• Jing-Ping Z, Tian QB, Sakagami H, Kondo H, Endo S, Suzuki T. p55 protein is a member of PSD scaffold proteins in the rat brain and interacts with various PSD proteins. Brain Res Mol Brain Res;135(1-2):204-16. 2005-04-27 [PMID: 15857683] (WB, Rat)
• Josephs KA, Uchikado H, McComb RD, Bashir R, Wszolek Z, Swanson J, Matsumoto J, Shaw G, Dickson DW. Extending the clinicopathological spectrum of neurofilament inclusion disease. Acta Neuropathol;109(4):427-32. 2005-04-01 [PMID: 15754170] (IF/IHC, Human)
• Cairns NJ, Grossman M, Arnold SE, Burn DJ, Jaros E, Perry RH, Duyckaerts C, Stankoff B, Pillon B, Skullerud K, Cruz-Sanchez FF, Bigio EH, Mackenzie IR, Gearing M, Juncos JL, Glass JD, Yokoo H, Nakazato Y, Mosaheb S, Thorpe JR, Uryu K, Lee VM, Trojanowski JQ. Clinical and neuropathologic variation in neuronal intermediate filament inclusion disease. Neurology;63(8):1376-84. 2004-10-26 [PMID: 15505152] (IF/IHC, Human)
• Rajasalu T, Teesalu K, Janmey PA, Uibo R. Demonstration of natural autoantibodies against the neurofilament protein alpha-internexin in sera of patients with endocrine autoimmunity and healthy individuals. Immunol Lett;94(1-2):153-60. 2004-06-15 [PMID: 15234547] (WB, Rat)
• Cairns NJ, Uryu K, Bigio EH, Mackenzie IR, Gearing M, Duyckaerts C, Yokoo H, Nakazato Y, Jaros E, Perry RH, Arnold SE, Lee VM, Trojanowski JQ. alpha-Internexin aggregates are abundant in neuronal intermediate filament inclusion disease (NIFID) but rare in other neurodegenerative diseases. Acta Neuropathol;108(3):213-23. 2004-09-01 [PMID: 15170578] (IF/IHC, Human)
• Cairns NJ, Zhukareva V, Uryu K, Zhang B, Bigio E, Mackenzie IR, Gearing M, Duyckaerts C, Yokoo H, Nakazato Y, Jaros E, Perry RH, Lee VM, Trojanowski JQ. alpha-internexin is present in the pathological inclusions of neuronal intermediate filament inclusion disease. Am J Pathol;164(6):2153-61. 2004-06-01 [PMID: 15161649] (IF/IHC, Human)
• Yuan A, Rao MV, Kumar A, Julien JP, Nixon RA. Neurofilament transport in vivo minimally requires hetero-oligomer formation. J Neurosci. 23(32):10463-5. 2003-11-12 [PMID: 14561875] (WB, Mouse)
• Ortino B, Inverardi F, Morante-Oria J, Fairen A, Frassoni C. Substrates and routes of migration of early generated neurons in the developing rat thalamus. Eur J Neurosci;18(2):323-32. 2003-07-01 [PMID: 12887414] (ICC/IF, Rat)
• Donner LR, Teshima I. Peripheral medulloepithelioma: an immunohistochemical, ultrastructural, and cytogenetic study of a rare, chemotherapy-sensitive, pediatric tumor. Am J Surg Pathol;27(7):1008-12. 2003-07-01 [PMID: 12826895] (IF/IHC, Human)
• Stachowiak EK, Fang X, Myers J, Dunham S, Stachowiak MK. cAMP-induced differentiation of human neuronal progenitor cells is mediated by nuclear fibroblast growth factor receptor-1 (FGFR1). J Neurochem;84(6):1296-312. 2003-03-01 [PMID: 12614330] (ICC/IF, Human)
• Talbot NC, Powell AM, Garrett WM. Spontaneous differentiation of porcine and bovine embryonic stem cells (epiblast) into astrocytes or neurons. In Vitro Cell Dev Biol Anim;38(4):191-7. 2002-04-01 [PMID: 12197769] (ICC/IF, Bovine, Porcine)
• McGraw TS, Mickle JP, Shaw G, Streit WJ. Axonally transported peripheral signals regulate alpha-internexin expression in regenerating motoneurons. J Neurosci. 2002-06-15 [PMID: 12077192] (IF/IHC, Rat)
• Lariviere RC, Nguyen MD, Ribeiro-da-Silva A, Julien JP. Reduced number of unmyelinated sensory axons in peripherin null mice. J Neurochem;81(3):525-32. 2002-05-01 [PMID: 12065660] (ICC/IF, WB, Mouse)
• Shaw G, Morse S, Ararat M, Graham FL. Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells. FASEB J. 2002-06-01 [PMID: 11967234] (ICC/IF, Human)
• Evans J, Sumners C, Moore J et al. Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem. J Neurophysiol. 2002-02-01 [PMID: 11826071] (ICC/IF, Rat)

FAQs for alpha-Internexin Antibody (NB300-140). (Showing 1 - 1 of 1 FAQs).

1. One of our customers bought the item NB300-140 (lot 9811) in November 2011. In February 2012, the ab was OK, but in March 2012 the antibody was coagulated in the vial. I think it's a contamination of the vial by any cause after opening, and we have no inquiry about a such observation by other customers. Did you have any inquiry about this antibody?
   • I had a look at our stock, and it appears to be fine, though I did notice some precipitate in the bottom of the tubes. I am not sure what this is at this stage, but will run some gels to find out. It should not be cell debris, since the cells are not in the same chamber as the antibody rich supernatant in the CL350 flasks. It may be aggregates of antibody molecules and/or serum proteins. Those tissue culture supernatants can contain very large amounts of antibody, so it would not be too surprising if some antibody precipitated out. I would recommend that your customer centrifuge that material out in the meantime and use the supernatant for their experiments.

Bioinformatics

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