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ECM220 Sigma-AldrichQCM 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.

SDS
References
Brochures
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Overview

Replacement Information

Key Spec Table

Detection Methods
Colorimetric Fluorescent

Description
Catalogue Number	ECM220
Trade Name	QCM Chemicon
Description	QCM Laminin Migration Assay (24-well, colorimetric)
Overview	Also 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 Delivered	1. 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 Ca²⁺) 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. CO₂incubator 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 method	Colorimetric Fluorescent
Quality Level	MQ100

Applications
Application	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.
Application Notes	The 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.

Applications

Application

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.

Application Notes

The 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 Conditions	ECM220 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 Size	1 kit
Material Package	Sufficient 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 overview	Pub Med ID
Wound healing--aiming for perfect skin regeneration. Martin, P Science, 276: 75-81 (1997) 1997 Show Abstract The healing of an adult skin wound is a complex process requiring the collaborative efforts of many different tissues and cell lineages. The behavior of each of the contributing cell types during the phases of proliferation, migration, matrix synthesis, and contraction, as well as the growth factor and matrix signals present at a wound site, are now roughly understood. Details of how these signals control wound cell activities are beginning to emerge, and studies of healing in embryos have begun to show how the normal adult repair process might be readjusted to make it less like patching up and more like regeneration.	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 During early granulation tissue formation of wound repair, new capillaries invade the fibrin clot, a process that undoubtedly requires an interaction of vascular cells with the wound provisional matrix composed mainly of fibrin, fibronectin, and vitronectin. Integrin alphaVbeta3 is the vascular cell receptor for these wound-associated adhesive proteins. Therefore, we investigated the expression of this receptor on new capillaries of healing full-thickness cutaneous porcine wounds. During granulation tissue formation, alphaVbeta3 was expressed specifically on capillary sprouts invading the central fibrin clot whereas the closely related integrin alphaVbeta5 failed to localize to these cells. Cyclic peptides or antibody antagonists of alphaVbeta3 specifically inhibited granulation tissue formation in a transient manner during the period of invasive angiogenesis. Immunolocalization studies revealed that alphaVbeta3 became aggregated and lost from sprouting vessels after treatment with a peptide antagonist. In contrast, beta 1 integrins were not modulated by this treatment. Once granulation tissue filled the wound and invasive angiogenesis terminated, the alphaVbeta3 showed little or no expression in the granulation tissue microvasculature. These data demonstrate that integrin alphaVbeta3 plays a fundamental, but transient, role during invasive angiogenesis and granulation tissue formation in a healing wound.	8623913

Reference overview

Pub Med ID

Brochure

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

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Title
QCM™ Laminin Migration Assay

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ECM220 Sigma-AldrichQCM 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.

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

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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.

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

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