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ECM510 QCM Chemotaxis Cell Migration Assay, 96-well (8 µm), fluorimetric

ECM510
1 plate  96 wells
Purchase on Sigma-Aldrich

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Tabulka spec. kláve

Key ApplicationsDetection Methods
ACTFluorescent
Description
Catalogue NumberECM510
Brand Family Chemicon®
Trade Name
  • QCM
  • Chemicon
DescriptionQCM Chemotaxis Cell Migration Assay, 96-well (8 µm), fluorimetric
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. Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely accepted of which is the Boyden Chamber assay. However, current methods of analysis are time-consuming and tedious, involving cotton swabbing of non-migrated cells on the top side of insert, manual staining and counting. Recently a fluorescence blocking membrane insert was introduced to address these issues; however, this approach requires labeling of the cells with Calcein-AM and extensive washing to remove free Calcein before cell migration. The effect of this treatment on cell behavior/migration remains questionable.

The Chemicon QCM™ 96-well Migration Assay does not require cell labeling, scraping, washing or counting. The 96-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples.

In the Chemicon QCM™ 96-well Migration Assay, migratory cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer. These cells are subsequently lysed and detected by the patented CyQuant GR dye (Molecular Probes). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids.

The Chemicon QCM™ 96-well Migration Assay provides a quick and efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for cell migration, or analysis of gene function in transfected cells.

The Chemicon QCM™ 96-well Migration Assay utilizes an 8 μm pore size, as this is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells; however, it is not appropriate for lymphocyte migration experiments. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.

In addition, Chemicon also provides QCM™ 24-well insert cell migration assay systems, CytoMatrix™ Cell Adhesion strips coated with ECM proteins or anti integrin antibodies, and QuantiMatrix™ ECM protein ELISA kits.

Application:

The Chemicon QCM™ 96-well Migration Assay is ideal for the study of chemotaxis cell migration. The quantitative nature of this assay is especially useful for large scale screening of pharmacological agents. The 8 μm pore size of this assay's Boyden chambers is appropriate for migration studies of most cell types. Each kit provides sufficient materials for the evaluation of 96 samples.

The Chemicon QCM™ 96-well Migration Assay is intended for research use only; not for diagnostic applications.
Materials Required but Not Delivered1. Precision pipettes: sufficient for aliquoting cells.

2. Harvesting buffer: EDTA or trypsin cell detachment buffer. Suggested formulations include a) 2 mM EDTA/PBS, b) 0.05% trypsin in Hanks Balanced Salt Solution (HBSS) containing 25 mM HEPES, or other cell detachment formulations as optimized by individual investigators.

Note: Trypsin cell detachment buffer maybe required for difficult cell lines. Allow sufficient time for cell receptor recovery.

3. Tissue culture growth medium appropriate for subject cells, such as DMEM containing 10% FBS.

4. Chemoattractants (eg. 10% FBS) or pharmacological agents for addition to culture medium, if screening is desired.

5. Quenching Medium: serum-free medium, such as DMEM, EMEM, or FBM (fibroblast basal media), containing 5% BSA.

Note: Quenching Medium must contain divalent cations (Mg2+, Ca2+) sufficient for quenching EDTA in the harvesting buffer.

6. Sterile PBS 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. Fluorescence plate reader.

13. Sterile cell culture hood.
Product Information
Components
  • Sterile 96-well Cell Migration Plate Assembly: (Part No. 90128) One 96-well feeder tray, and one 96-well Cell Migration Chamber plate. Area = 0.3 cm2, 50,000 cells per well
  • 96-well Cell Culture Tray: (Part No. 90129) One 96-well feeder tray.
  • Cell Detachment Solution: (Part No. 90131) One bottle - 16 mL.
  • 4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL.
  • CyQuant GR Dye®: (Part No. 90132) One vial - 75 μL
Detection methodFluorescent
Quality LevelMQ100
Applications
ApplicationThe QCM 8 uM 96-well Migration Assay utilizes a 8 um pore size, which is appropriate for leukocyte migration.
Key Applications
  • Activity Assay
Biological Information
Species Reactivity
  • All
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.
Packaging Information
Material Size1 plate
Material Package96 wells
Global Trade Item Number
Katalogové číslo GTIN
ECM510 04053252506604