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QCM™ 24-well Fluorimetric Cell Migration Assay Kit

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号:ECM509

牌:Millipore 密理博

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Replacement Information

重要规格表

Key Applications Detection Methods
ACT Fluorescent
Description
Catalogue Number ECM509
Brand Family Chemicon®
Trade Name
  • QCM
  • Chemicon
Description QCM Chemotaxis Cell Migration Assay, 24-well (8 µm), fluorimetric
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. 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™ 24-well Cell Migration Assay does not require cell labeling, scraping, washing or counting. The 24-well insert and homogenous fluorescence detection format allows for large-scale screening and quantitative comparison of multiple samples.

In the CHEMICON® QCM™ 24-well Migration Assay, invaded 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 CyQUANTâ GR dye (Molecular Probes) (1-2). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (3).

The CHEMICON® QCM™ 24-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™ 24-well Migration Assay utilizes an 8 mm 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 continues to provide numerous migration, invasion, and adhesion products including:

· QCM™ 8μm 96-well Chemotaxis Cell Migration Assay (ECM510)

· QCM™ 5μm 96-well Chemotaxis Cell Migration Assay (ECM512)

· QCM™ 3μm 96-well Chemotaxis Cell Migration Assay (ECM515)

· QCM™ 96-well Cell Invasion Assay (ECM555)

· QCM™ 96-well Collagen-based Cell Invasion Assay (ECM556)

· 24-well Insert Cell Migration and Invasion Assay Systems

· CytoMatrix™ Cell Adhesion strips (ECM protein coated)

· QuantiMatrix™ ECM protein ELISA kits

Test Principle

The CHEMICON® QCM™ 24-well Cell Migration Assay is performed in a Migration Chamber, based on the Boyden chamber principle. Each kit contains 24 inserts; each insert utilizes an 8 mm pore size polycarbonate membrane, 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.
Materials Required but Not Delivered 1. 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.
References
Product Information
Components
  • Sterile 24-well Cell Migration Plate Assembly: (Part No. 90333) Two 24-well plates with 12 inserts per plate (24 inserts total/kit).
  • Cell Detachment Solution: (Part No. 90131) One bottle - 16 mL.
  • 4X Cell Lysis Buffer: (Part No. 90130) One bottle - 16 mL.
  • CyQUANTâ GR Dye1: (Part No. 90132) One vial - 75 μL
  • Forceps: (Part No. 10203) One each.
Detection method Fluorescent
Applications
Application The QCM 24-well Migration Assay is ideal for the study of chemotaxis cell migration. The assay uses a 24-well plate with an 8 micron pore size, with fluorescent detection.
Key Applications
  • Activity Assay
Application Notes The CHEMICON® QCM™ 24-well Migration Assay is ideal for the study of chemotaxis cell migration. Each Chemicon Cell Migration Assay Kit contains sufficient reagents for the evaluation of 24 samples. The quantitative nature of this assay is especially useful for screening of pharmacological agents.

The CHEMICON® QCM™ 24-well Migration Assay is intended for research use only; not for diagnostic applications.
Biological Information
Species Reactivity
  • All
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 Store kit materials at 2-8°C for up to their expiration date. Do not freeze.
Packaging Information
Material Size 1 plate
Material Package 24 wells
Transport Information
Supplemental Information
Specifications

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