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ECM220 QCM Laminin Migration Assay (24-well, colorimetric)

This QCM Laminin Migration Assay incorporates a 5 um porous membrane, 24-well plate Boyden chamber & mouse laminin, a major extracellular matrix (ECM) protein in basement membrane, to examine cell migration dependent upon the presence of laminin.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
ECM220
1 kit  Sufficient for 24 samples
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

Detection Methods
Colorimetric Fluorescent
Description
Catalogue NumberECM220
Trade Name
  • QCM
  • Chemicon
DescriptionQCM Laminin Migration Assay (24-well, colorimetric)
OverviewAlso available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here

Introduction
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation (1, 2).
Cell migration may be evaluated using several different methods; the most widely accepted being the Boyden Chamber assay. The Boyden Chamber system uses a two-chamber plate model in which a porous membrane provides an interface between two chambers. Cells are seeded in the upper chamber and chemoattractants placed in the lower chamber. Cells in the upper chamber migrate toward the chemoattractants by passing through the porous membrane to the lower chamber. Migratory cells are stained and quantified. Most cells are sized from 30 to 50 µm can migrate through 3 to 10 µm pore.
Materials Required but Not Delivered1. Precision pipettes, sufficient for aliquoting appropriate volume of cells and reagents.
2. Harvesting buffer: EDTA or trypsin-based cell detachment buffer, or other cell detachment formulations as optimized by individual investigators. Millipore’s ready-to-use non-mammalian detachment solution, Accutase (Cat. No. SCR005) is recommended.
Note: Trypsin-based cell detachment buffer may be required for strongly adherent cell lines, but can strip cell surface proteins. Allow sufficient time for cell receptor recovery.

3. Tissue culture growth medium appropriate for subject cells.
4. Quenching Buffer: Serum-free medium such as DMEM or RPMI-1640, containing 5% BSA
Note: Quenching Buffer must contain sufficient divalent cations (Mg 2+ or Ca 2+) to quench any EDTA present in the Harvesting Buffer.

5. Chemoattractants (eg. 10% FBS) or pharmacological agents for addition to culture medium, if screening is desired.
6. Sterile PBS (Cat. No. BSS-1005-B) or HBSS to wash cells.
7. Distilled water.
8. Low speed centrifuge and tubes for cell harvesting.
9. CO2 incubator appropriate for subject cells.
10. Hemocytometer or other means of counting cells.
11. Trypan blue or equivalent viability stain.
12. Spectrophotometer (microplate reader with 540-570 nm detection capability).
13. Sterile cell culture hood.
14. (Optional) Graduated ocular (calibrated), or automated method for counting stained cells on a membrane.
References
Product Information
Components
  • ECM220-1
  • 1. 24-well Cell Migration Plate Assembly - 5 µm: (Part No. 2005708) Two 24-well plates, each containing 12- 5 µm pore inserts per plate.
  • 2. BSA 30% solution: (Part No. CS203352) One vial containing 0.5 mL of a 30% solution.
  • 3. Cell Stain Solution*: (Part No. 90144) One bottle.
  • 4. Extraction Buffer: (Part No. 90145) One bottle.
  • 5. 24-well Stain Receiver Plate: (Part No. CR209941) One each.
  • 6. 96-Well Stain Quantitation Plate: (Part No. 2005870) One each.
  • 7. Swabs: 50 ea
  • 8. Forceps: 1 pair
  • *Caution: Cell Stain Solution contains a small amount of crystal violet, which is toxic if swallowed or inhaled, and may cause irritation to the eyes, respiratory system, and skin. Handle with caution.
  • ECM220-2
  • 1. Laminin: (Part No. CS202685) Two vials each containing 60 μg in solution.
Detection methodColorimetric Fluorescent
Quality LevelMQ100
Applications
ApplicationThis QCM Laminin Migration Assay incorporates a 5 um porous membrane, 24-well plate Boyden chamber & mouse laminin, a major extracellular matrix (ECM) protein in basement membrane, to examine cell migration dependent upon the presence of laminin.
Application NotesThe Millipore QCM Laminin Migration Assay incorporates a 5 µm porous membrane Boyden chamber and mouse laminin, a major extracellular matrix (ECM) protein in basement membrane, to examine cell migration dependent upon the presence of laminin. The laminin is isolated from mouse sarcoma and it is enriched with mouse laminin 1 (laminin-111). This ECM promotes cell adhesion, proliferation as well as migration in many cell types such as embryonic stem cells, neural cells, liver cells and cancer cells. Cells interact with laminin through integrins α1β1, α2β2, α3β1, α6β1, α7β1, and α6β4. The assay is designed specifically to examine the migration of a subset of fibroblasts, cancer cells and neural cells.
Laminin protein is coated on the bottom of each chamber’s porous membrane as the driver for cell migration. A similarly BSA-coated control chamber(s) provides an appropriate migration control. Cells that migrate toward laminin are stained using crystal violet, a nucleic dye, extracted from the membrane surface, and quantified on a microplate reader using spectrophotometry.

Each kit provides sufficient materials for the evaluation of 24 samples.
Biological Information
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 ConditionsECM220 should be used within four months from date of receipt. In that time:
• ECM220-1 kit components should be stored at 2° to 8°C. DO NOT FREEZE.
• ECM220-2 component must be stored at -20°C until needed.
Packaging Information
Material Size1 kit
Material PackageSufficient for 24 samples
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
ECM220 04053252014635

Documentation

QCM Laminin Migration Assay (24-well, colorimetric) SDS

Title

Safety Data Sheet (SDS) 

References

Reference overviewPub Med ID
Wound healing--aiming for perfect skin regeneration.
Martin, P
Science, 276: 75-81 (1997)  1997

Show Abstract 		9082989 9082989
Transient functional expression of alphaVbeta 3 on vascular cells during wound repair.
Clark, R A, et al.
Am. J. Pathol., 148: 1407-21 (1996)  1996

Show Abstract 		8623913 8623913

User Guides

Title
QCM™ Laminin Migration Assay

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