ECM221 | QCM Laminin Migration Assay (24-well, fluorometric)

ECM221
1 kit  Sufficient for 12 assays
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      Overview

      Replacement Information

      Key Spec Table

      Detection Methods
      Fluorescent
      Description
      Catalogue NumberECM221
      Trade Name
      • QCM
      • Chemicon
      DescriptionQCM Laminin Migration Assay (24-well, fluorometric)
      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.

      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. Fluorescent microplate reader with 540-570 nm detection capability (FITC channel).
      13. Sterile cell culture hood.
      References
      Product Information
      Components
      • ECM221-1
      • 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. 24-well Stain Receiver Plate: (Part No. PIMWS2450) One each.
      • 4. 96-Well Black Quantitation Plate: (Part No. CR201763) One each.
      • 5. Forceps: 1 pair
      • ECM221-2
      • 1. Laminin: (Part No. CS202685) Two vials each containing 60 μg in solution.
      • 2. Accutase™: (Cat. No. SCR005) One 100 mL bottle of cell detachment solution.
      • 3. Calcein-AM: (Part No. CS202541) One 50 µg vial.
      Detection methodFluorescent
      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 Calcein-AM™ a living cell fluorescent dye, quantified on a fluorescent microplate reader.

      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 ConditionsECM221 should be used within four months from date of receipt. In that time:
      • ECM221-1 kit components should be stored at 2° to 8°C. DO NOT FREEZE.
      • ECM221-2 kit components must be stored at -20°C until needed.
      Packaging Information
      Material Size1 kit
      Material PackageSufficient for 12 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      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

      Brochure

      Title
      Advancing cancer research: From hallmarks & biomarkers to tumor microenvironment progression
      Cell Migration and Invasion: Choosing the Right Assay

      User Guides

      Title
      QCM™ Laminin Migration Assay

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      Categories

      Life Science Research > Cell Analysis > Cell-based Assays > Cell Migration Assays