Neurite Outgrowth Assay Kit (1µM)

Detection Methods
Chromogenic

Detection Methods

Chromogenic

Description
Catalogue Number	NS225
Brand Family	Chemicon®
Trade Name	Chemicon
Description	Neurite Outgrowth Assay Kit (1 µm)
Materials Required but Not Delivered	1. 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 Information	Application 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.

Description

Catalogue Number

NS225

Brand Family

Chemicon®

Trade Name

Chemicon

Description

Neurite Outgrowth Assay Kit (1 µm)

Materials Required but Not Delivered

1. 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 Information

Application 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.

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 method	Chromogenic

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 method

Chromogenic

Applications
Application	The 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.

Applications

Application

The 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

Biological Information

Species Reactivity

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.

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 Conditions	When 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.

Storage and Shipping Information

Storage Conditions

When 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 Size	12 assays

Packaging Information

Material Size

12 assays

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

Neurite Outgrowth Assay Kit (1µM)

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