NS225 Neurite Outgrowth Assay Kit (1 µm)

NS225
12 assays  
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      Overview

      Replacement Information

      Key Specifications Table

      Detection Methods
      Chromogenic
      Description
      Catalogue NumberNS225
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionNeurite Outgrowth Assay Kit (1 µm)
      Materials Required but Not Delivered1. Desired extracellular matrix membrane coating protein, e.g., bovine collagen I or mouse laminin.
      2. Tissue culture instruments/supplies (including 37°C incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
      3. Differentiation media/factors for cell type of interest (e.g., nerve growth factor, serum-free media).
      4. 1X phosphate buffered saline (PBS) with Ca2+/Mg2+.
      5. Inverted microscope for neurite visualization on membrane inserts.
      6. Parafilm and spectrophotometer for neurite stain extraction and quantification.
      7. Protein lysis buffer (optional)
      Background InformationApplication CHEMICON's NS225 Neurite Outgrowth Assay Kit (1 μm) utilizes microporous tissue culture insert technology from Millipore and is based on the use of Millicell cell culture inserts (chambers) containing a permeable membrane with 1 μm pores at the base. The inserts are designed to fit into a receiver vessel, which when in use contains differentiation media in contact with the bottom of the insert membrane. The permeable membranes allow for discrimination between neurites and cell bodies, as projecting neurites will pass easily through the pores but cell bodies will not. Therefore, by inducing neurites to traverse these membrane pores, a purified population of neurites is located on the underside of the membrane, distal to the surface on which neural cell bodies are deposited. This kit has has been specifically designed for and validated with PC12 cells. The diameter of PC12 cell bodies is relatively small compared to that of many other cell types commonly used in neurite outgrowth assays [5, 6, 7]. As a result, many PC12 cell bodies may pass though pores larger than 1 μm, significantly obscuring neurite outgrowth assay results. Using a 1 μm pore size greatly reduces background and improves the signal-to-noise ratio in studies using PC12 cells. Use of this 1 μm pore size with larger cell types that extend wider diameter neurites, e.g., N1E-115 cells, will greatly diminish the number of neurites able to project through to the underside of the membrane, and is thus not recommended. The assay is a simple, efficient, and versatile system for the quantitative determination of compounds that influence neurite formation and repulsion. With this system, it is possible to screen numerous biological and pharmacological agents simultaneously, directly evaluate adhesion and guidance receptor functions responsible for neurite extension and repulsion, as well as analyze gene function in transfected cells. The microporous filter allows for biochemical separation and purification of neurites and cell bodies for detailed molecular analysis of protein expression, signal transduction processes and identification of drug targets that regulate neurite outgrowth or retraction processes. Reagents and materials supplied in the NS225 Neurite Outgrowth Assay Kit are sufficient for 12 tests.
      References
      Product Information
      Components
      • Neurite Outgrowth Plate Assembly, 1 µm: (Part No. 2007254) 1 x 24-well plate containing 12 x Millicell hanging inserts with 1 µm pore size membranes.
      • Neurite Stain Solution: (Part No. 90242) One 20 mL bottle.
      • Neurite Stain Extraction Buffer: (Part No. 90243) One 20 mL bottle.
      • Neurite Outgrowth Assay Plate: (Part No. 2007255) Two 24-well plates.
      • Cotton Swabs: (Part No. 10202) 50 swabs.
      • Forceps: (Part No. 10203) One each.
      Detection methodChromogenic
      Applications
      ApplicationThe NS225 Neurite Outgrowth Assay Kit (1 μm) is based on the use of Millicell cell culture inserts (chambers) containing a permeable membrane with 1 μm pores at the base.
      Biological Information
      Species Reactivity
      • All
      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 ConditionsWhen stored at 2-8° C, the kit components are stable up to the expiration date. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date.
      Packaging Information
      Material Size12 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Neurite Outgrowth Assay Kit (1 µm) SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Neural progenitor cells derived from adult bone marrow mesenchymal stem cells promote neuronal regeneration.
      Yue Tang,Yong-Chun Cui,Xiao-Juan Wang,Ai-Li Wu,Guang-Fu Hu,Fu-Liang Luo,Jia-Kang Sun,Jing Sun,Li-Ke Wu
      Life sciences  91  2012

      Show Abstract
      23000028 23000028
      Small GTPase Tc10 and its homologue RhoT induce N-WASP-mediated long process formation and neurite outgrowth.
      Abe, Tomoyuki, et al.
      J. Cell. Sci., 116: 155-68 (2003)  2003

      Show Abstract
      12456725 12456725
      Spinal cord repair: strategies to promote axon regeneration.
      McKerracher, L
      Neurobiol. Dis., 8: 11-8 (2001)  2001

      Show Abstract
      11162236 11162236
      Repulsive factors and axon regeneration in the CNS
      Fournier, A E and Strittmatter, S M
      Curr Opin Neurobiol, 11:89-94 (2001)  2001

      11179877 11179877
      Glial inhibition of nerve regeneration in the mature mammalian CNS.
      Qiu, J, et al.
      Glia, 29: 166-74 (2000)  2000

      Show Abstract
      10625335 10625335
      Axon regeneration: Vaccinating against spinal cord injury
      Filbin, M T
      Curr Biol, 10:R100-R103 (2000)  2000

      10679313 10679313
      Extracellular matrix allows PC12 neurite elongation in the absence of microtubules
      Lamoureux, P, et al.
      J Cell Biol, 110:71-79 (1990)  1990

      2153148 2153148
      Nerve growth factor-induced neurite outgrowth in PC12 cells involves the coordinate induction of microtubule assembly and assembly-promoting factors
      Drubin, D G, et al
      J Cell Biol, 101:1799-1807 (1985)  1985

      2997236 2997236

      Data Sheet

      Title
      Neurite Outgrowth Assay Kit (1um) - Data Sheet

      Posters

      Title
      Poster: High Throughput Screening To Identify Novel Molecular Pathways Involved In Neurite Outgrowth

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      Categories

      Life Science Research > Cell Analysis > Cell-based Assays > Neurite Outgrowth