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ECM557 Sigma-AldrichQCM™ Leukocyte Transendothelial Migration Assay (Colorimetric, 24 Assays)

This Colorimetric QCM Leukocyte Transendothelial Cell Migration Assay provides an efficient model to analyze the ability of leukocytes to migrate through the endothelium.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

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ECM557

1 kit 24 Assays

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Key Specifications Table

Detection Methods
Colorimetric

Description
Catalogue Number	ECM557
Trade Name	QCM Chemicon
Description	QCM™ Leukocyte Transendothelial Migration Assay (Colorimetric, 24 Assays)
Overview	Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here Introduction Millipore’s Colorimetric QCM™ Leukocyte Transendothelial Cell Migration Assay provides an efficient model to analyze the ability of leukocytes to migrate through the endothelium. The assay is designed with a 3 µm pore size cell culture insert, appropriate for most leukocyte migration. The upper side of the cell culture insert is coated with fibronectin to support the optimal attachment and growth of endothelial cells. The assay allows investigators to evaluate the effects of various factors that influence the transendothelial process. Precoated cell culture inserts are provided in the Millipore QCM Leukocyte Transendothelial Cell Migration Assay to significantly reduce assay time. Additionally, the assay allows quantitative analysis of leukocyte migration. Following incubation of leukocytes with the endothelial cell layer, migratory cells are stained and quantified with WST-1 reagent and measured using a spectrophotometer. Absorbance correlates with cell migration.
Materials Required but Not Delivered	1. Endothelial cells (for example: HUVECs, Cat No. SCCE001) 2. Endothelial cell culture medium (Cat. No. SCME001) 3. Leukocytes and cell culture medium 4. Harvesting buffer: Millipore’s cell detachment solution, Accutase™ (Cat. No. SCR005), EDTA or trypsin-based cell detachment buffer, or other cell detachment formulations as optimized by individual investigators can also be used. 5. Serum-free medium, such as RPMI-1640, DMEM (Cat No. ES-101-B), MEM, etc. containing 0.5% BSA (Cat No. 82-046-4), Pen/Strep (Cat No. TMS-AB2-C). 6. Sterile PBS (Cat No. BSS-1006-B) to wash cells. 7. Distilled water (Cat No. TMS-006-B) 8. (Optional) Chemoattractant or pharmacological agent for addition to culture medium. 9. Low speed centrifuge and tubes for cell harvesting. 10. CO2 incubator appropriate for subject cells. 11. Microplate reader (420 - 480 nm detection) or a spectrophotometer.
Background Information	Leukocytes patrol the vascular system. They must migrate across endothelial barriers in order to recruit to sites of inflammation. This process involves a multistep cascade consisting of leukocyte rolling, adhesion, and transmigration. A quantitative assay for leukocyte transendothelial migration has been described using a modified Boyden chamber system (Roth et al. 1995, Ding et al. 2000). The Boyden chamber system is a two chamber system with a porous membrane providing an interface between these two chambers. Endothelial cells are cultured on top of the porous membrane that is coated with an extracelluar matrix (ECM) protein. Once a confluent layer of cells is established, leukocytes are then added onto the endothelial monolayer. Leukocyte migration across the endothelium is determined by measuring the number of cells that migrate between the endothelial cells, through the porous membrane, to the lower chamber.

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Product Information
Components	1. Migration Test Plate: (Part No. CS202626) Two 24-well culture plates, each containing 12 human fibronectin-coated cell culture inserts (3 μm pore size), sufficient for the evaluation of 24 test samples 2. TNFα: (Part No. 2004167) One tube - 20 μL at 0.1 mg/mL 3. Cell Stain Solution: (Part No. 20294) One vial - 10 mL 4. WST-1 Reagent: (Part No. CS201370) One vial. 5. Electro Coupling Solution: (Part No. CS201382) One vial - 5 mL. 6. 96 well Stain Quantitation Plate: (Part No. 2005870) One plate. 7. Forceps: (Part No. 10203) 1 pair
Detection method	Colorimetric
Quality Level	MQ100

Applications
Application	This Colorimetric QCM Leukocyte Transendothelial Cell Migration Assay provides an efficient model to analyze the ability of leukocytes to migrate through the endothelium.
Application Notes	Results of the assay may be illustrated graphically using a bar chart to display optical density (OD) values at 420 - 480 nm. Additional migration may also be induced or inhibited in test wells through the addition of cytokines or other pharmacological agents. Figures 2 and 3 demonstrate typical migration results. One should use the data for reference only. This data should not be used to interpret actual assay results.

Applications

Application

This Colorimetric QCM Leukocyte Transendothelial Cell Migration Assay provides an efficient model to analyze the ability of leukocytes to migrate through the endothelium.

Application Notes

Results of the assay may be illustrated graphically using a bar chart to display optical density (OD) values at 420 - 480 nm.

Additional migration may also be induced or inhibited in test wells through the addition of cytokines or other pharmacological agents.
Figures 2 and 3 demonstrate typical migration results. One should use the data for reference only. This data should not be used to interpret actual assay results.

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	The kit is packaged in two individual boxes. ECM557-1 contains components that upon arrival should be stored at 2° to 8°C. ECM557-2 contains TNFα , WST-1, and Electro Coupling Solution that upon arrival should be stored at -20°C. Please refer to kit label for expiration date.

Packaging Information
Material Size	1 kit
Material Package	24 Assays

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalog Number	GTIN
ECM557	04053252004674

Documentation

QCM™ Leukocyte Transendothelial Migration Assay (Colorimetric, 24 Assays) SDS

Title
Safety Data Sheet (SDS)

References

Reference overview	Pub Med ID
Regulation of chemokine-induced transendothelial migration of T lymphocytes by endothelial activation: differential effects on naive and memory T cells. Ding, Z, et al. J. Leukoc. Biol., 67: 825-33 (2000) 2000	10857855
Characterization of transendothelial chemotaxis of T lymphocytes. Roth, S J, et al. J. Immunol. Methods, 188: 97-116 (1995) 1995	8551044

User Guides

Title
QCM Leukocyte Transendothelial Migration Assay - Colorimetric

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ECM557 Sigma-AldrichQCM™ Leukocyte Transendothelial Migration Assay (Colorimetric, 24 Assays)

This Colorimetric QCM Leukocyte Transendothelial Cell Migration Assay provides an efficient model to analyze the ability of leukocytes to migrate through the endothelium.

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