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MAB5266 Anti-Neurofilament 200 kDa Antibody, clone N52

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MAB5266
50 µg  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, M, Mk, Po, RWB, IHCMPurifiedMonoclonal Antibody
      Description
      Catalogue NumberMAB5266
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Neurofilament 200 kDa Antibody, clone N52
      Background InformationNeurofilaments are a type of intermediate filament that serve as major elements of the cytoskeleton supporting the axon cytoplasm. They are the most abundant fibrillar components of the axon, being on average 3-10 times more frequent than axonal microtubules. Neurofilaments (10nm in dia.) are built from three intertwined protofibrils which are themselves composed of two tetrameric protofilament complexs of monomeric proteins. The neurofilament triplet proteins (68/70, 160, and 200 kDa) occur in both the central and peripheral nervous system and are usually neuron specific. The 68/70 kDa NF-L protein can self-assemble into a filamentous structure, however the 160 kDa NF-M and 200 kDa NF-H proteins require the presence of the 68/70 kDa NF-L protein to co-assemble. Neuromas, ganglioneuromas, gangliogliomas, ganglioneuroblastomas and neuroblastomas stain positively for neurofilaments. Although typically restricted to neurons, neurofilaments have been detected in paragangliomas and adrenal and extra-adrenal pheochromocytomas. Carcinoids, neuroendocrine carcinomas of the skin, and oat cell carcinomas of the lung also express neurofilaments. For more neurofilament information see Nervous System Cell Type Specific Marker chart online under the CHEMICON Technical Support section.
      References
      Product Information
      FormatPurified
      PresentationPurified immunoglobulin (Ion exchange chromatography). Liquid in 0.02M phosphate buffer, 0.25M NaCl with 0.1% sodium azide, pH 7.6
      Quality LevelMQ100
      Applications
      ApplicationThis Anti-Neurofilament 200 kDa Antibody, clone N52 is validated for use in WB, IH for the detection of Neurofilament 200 kDa.
      Key Applications
      • Western Blotting
      • Immunohistochemistry
      Application NotesWestern blot: 5-10 μg/mL

      Immunohistochemistry: 5-10 μg/mL

      Optimal working dilutions must be determined by end user.

      Immunohistochemistry: Antibody N52 reacts with a fragment of NF-200 side-arm that is located at the end of the MPR KSP domain and which contains the consensus cdk-5 phosphorylation site, however this reactivity is abolished if the NF-200 fragment becomes phosphorylated by cdk-5 {Guidato et al, 1996}. Thus for the fullest staining and reactivity, including westerns, it is suggested that samples be treated with alkaline phosphatase prior to antibody staining. The following treatment protocol is suggested:



      ProtocolPreparation of Solutions:

      I. TBS: Tris-HCl, 0.1 mol/l; NaCl, 0.1 mol/l' pH 8.0.

      II. Mix alkaline phosphatase, 1 mg/ml with 1 mol/l ZnSO4,; 1mol/l MgCl2, and 1mmol/l PMSF; and dissove in TBS.

      NOTE: It is necessary to dialyze the alkaline phosphatase against solution I before use.

      Procedure:

      Frozen sections from shock frozen tissue samples are air-dried and then fixed with acetone for 10 min at -20°C. The excess acetone is allowed to evaporate at room temperature. Incubate the sections in solutions II for 4 hours at +30oC and wash in TBS 3 times for 5 minutes each. Negative control sections are incubated in solution II with out alkaline phosphatase. Further treatment as follows:

      - Cover the preparation with 10-20 μl antibody solution and incubate for 1 hr in a humid chamber.

      - Immerse the slide in TBS and wash in TBS 3 times for 5 minutes each.

      - Cover the preparation with 10-20 μl of a solution of anti-mouse Ig-antibody, labeled with fluorscine isothiocyanate, and incubate in a humid chamber at 37°C for 1 hour.

      - Wash the slide in TBS 3 times for 5 min each.
      Biological Information
      ImmunogenCarboxy terminal tail segment of enzymatically dephosphorylated porcine H-chain.
      CloneN52
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityMAB5266 reacts with the phophorylated and dephosphorylated H-chain of neurofilament 200kDa (NF-H) in normal tissues/extracts (Shaw, 1986). MAB5266 can be used to detect cells of neuronal origin by immunohistochemistry and Western blot. It has been reported that reactivity of MAB5266 with NF-H is blocked in cdk-5 over-expressing cells (Guidato, 1996).
      IsotypeIgG1
      Species Reactivity
      • Human
      • Mouse
      • Monkey
      • Pig
      • Rat
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • NEFH
      • NFH
      • NF-H
      • KIAA0845
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P12036 # Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins.
      SIZE: 1026 amino acids; 112480 Da
      PTM: There are a number of repeats of the tripeptide K-S-P, NFH is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFH results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber. & Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincident with a change in the neurofilament function.
      DISEASE: SwissProt: P12036 # Defects in NEFH are a cause of susceptibility to amyotrophic lateral sclerosis (ALS) [MIM:105400]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons, and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology is likely to be multifactorial, involving both genetic and environmental factors.
      SIMILARITY: SwissProt: P12036 ## Belongs to the intermediate filament family.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain at 2-8°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
      Packaging Information
      Material Size50 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-Neurofilament 200 kDa Antibody, clone N52 SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-Neurofilament 200 kDa Antibody, clone N52 Certificates of Analysis

      TitleLot Number
      MOUSE ANTI-NEUROFILAMENT 200KD - 3231555 3231555
      MOUSE ANTI-NEUROFILAMENT 200KD - 3439218 3439218
      MOUSE ANTI-NEUROFILAMENT 200KD - 3708741 3708741
      MOUSE ANTI-NEUROFILAMENT 200KD -2567008 2567008
      MOUSE ANTI-NEUROFILAMENT 200KD -2813877 2813877
      MOUSE ANTI-NEUROFILAMENT 200KD MONOCLONAL ANTIBODY 2986162
      MOUSE ANTI-NEUROFILAMENT 200KD MONOCLONAL ANTIBODY 2901542
      MOUSE ANTI-NEUROFILAMENT 200KD MONOCLONAL ANTIBODY - 2123144 2123144
      MOUSE ANTI-NEUROFILAMENT 200KD MONOCLONAL ANTIBODY - 2147058 2147058
      MOUSE ANTI-NEUROFILAMENT 200KD MONOCLONAL ANTIBODY - 2391078 2391078

      References

      Reference overviewApplicationSpeciesPub Med ID
      Selective conversion of fibroblasts into peripheral sensory neurons.
      Blanchard, JW; Eade, KT; Szűcs, A; Lo Sardo, V; Tsunemoto, RK; Williams, D; Sanna, PP; Baldwin, KK
      Nature neuroscience  18  25-35  2015

      Show Abstract
      25420069 25420069
      Neurotrophin-4 regulates the survival of gustatory neurons earlier in development using a different mechanism than brain-derived neurotrophic factor.
      Patel, AV; Krimm, RF
      Developmental biology  365  50-60  2012

      Show Abstract
      22353733 22353733
      Sensory-motor deficits and neurofilament disorganization in gigaxonin-null mice.
      Ganay, T; Boizot, A; Burrer, R; Chauvin, JP; Bomont, P
      Molecular neurodegeneration  6  25  2011

      Show Abstract Full Text Article
      21486449 21486449
      The BTB and CNC homology 1 (BACH1) target genes are involved in the oxidative stress response and in the control of the cell cycle
      Warnatz HJ, Schmidt D, Manke T, Piccini I, Sultan M, Borodina T, Balzereit D, Wruck W, Soldatov A, Vingron M, Lehrach H, Yaspo ML
      J Biol Chem  2011

      Show Abstract
      21555518 21555518
      BDNF is required for the survival of differentiated geniculate ganglion neurons.
      Patel, Ami V and Krimm, Robin F
      Dev. Biol., 340: 419-29 (2010)  2010

      Show Abstract
      20122917 20122917
      Contribution of voltage-gated sodium channels to the b-wave of the mammalian flash electroretinogram.
      Mojumder, DK; Sherry, DM; Frishman, LJ
      The Journal of physiology  586  2551-80  2008

      Show Abstract
      18388140 18388140
      Subcellular compartmentalization of two calcium binding proteins, calretinin and calbindin-28 kDa, in ganglion and amacrine cells of the rat retina.
      Mojumder, DK; Wensel, TG; Frishman, LJ
      Molecular vision  14  1600-13  2008

      Show Abstract
      18769561 18769561
      Aminoglycoside-induced degeneration of adult spiral ganglion neurons involves differential modulation of tyrosine kinase B and p75 neurotrophin receptor signaling.
      Justin Tan, Robert K Shepherd, Justin Tan, Robert K Shepherd, Justin Tan, Robert K Shepherd, Justin Tan, Robert K Shepherd
      The American journal of pathology  169  528-43  2006

      Show Abstract Full Text Article
      16877354 16877354
      Effects of spinal nerve ligation on immunohistochemically identified neurons in the L4 and L5 dorsal root ganglia of the rat.
      Hammond, Donna L, et al.
      J. Comp. Neurol., 475: 575-89 (2004)  2004

      15236238 15236238
      Adeno-associated viral transfer of opioid receptor gene to primary sensory neurons: a strategy to increase opioid antinociception.
      Xu, Y, et al.
      Proc. Natl. Acad. Sci. U.S.A., 100: 6204-9 (2003)  2003

      Show Abstract
      Immunoblotting (Western)Rat12719538 12719538

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