ECM555 QCM ECMatrix Cell Invasion Assay, 96-well (8 µm), fluorimetric

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ECM555
1 plate  96 wells
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      Overview

      Replacement Information

      Key Specifications Table

      Key ApplicationsDetection Methods
      ACT Fluorescent
      Description
      Catalogue NumberECM555
      Brand Family Chemicon®
      Trade Name
      • QCM
      • Chemicon
      DescriptionQCM ECMatrix Cell Invasion Assay, 96-well (8 µm), fluorimetric
      OverviewAlso available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here

      Introduction
      Invasion through the extracellular matrix (ECM) is an important step in tumor metastasis. Cancer cells initiate invasion by adhering to and spreading along the blood vessel wall. Proteolytic enzymes, such as MMP collagenases, dissolve tiny holes in the sheath-like covering (basement membrane) surrounding the blood vessels to allow cancer cells to invade (Egeblad & Werb, 2002).

      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-invaded cells on the topside 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 invasion. The effect of this treatment on cell behavior/invasion remains questionable.

      The CHEMICON QCM™ 96-well Invasion 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 Invasion 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 CyQuant GR* dye (Molecular Probes) (Gildea et al., 2000). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (Jones et al., 2001).

      *CyQUANT GR® is a registered trademark of Molecular Probes, Inc. The reagent is licensed from Molecular Probes, Inc. and is for use in kits sold by Chemicon International, Inc. for the monitoring of cell invasion and cell migration only.

      The CHEMICON Cell Invasion Assay Kit provides an efficient system for evaluating the invasion of tumor cells through a basement membrane model. The kit utilizes ECMatrixTM, a reconstituted basement membrane matrix of proteins derived from the Engelbreth Holm-Swarm (EHS) mouse tumor (Albini et al., 1987; Repesh, 1989; Terranova et al., 1986; Liotta, 1984). We examined the kit's performance using human fibrosarcoma (HT-1080) and non-invasive fibroblasts (NIH3T3).

      The CHEMICON QCM™ 96-well Invasion Assay provides a quick and efficient system for quantitative determination of various factors on cell invasion, including screening of pharmacological agents, evaluation of MMPs or other proteases responsible for cell invasion, or analysis of gene function in transfected cells.

      In addition, CHEMICON also provides QCM™ 96-well Chemotaxis Cell Migration Assay Systems (8 μm or 3 μm pore), 24-well insert Cell Migration and Invasion Assay Systems, CytoMatrix™ Cell Adhesion strips coated with ECM proteins or anti-integrin antibodies, and QuantiMatrix™ ECM protein ELISA kits.

      Test Principle:

      The CHEMICON Cell Invasion Assay is performed in a 96 well invasion plate based on the Boyden chamber principle. This plate contains 96 inserts; each insert contains an 8 μm pore size polycarbonate membrane coated with a thin layer of ECMatrixTM. The ECM layer occludes the membrane pores, blocking non-invasive cells from migrating through. Invasive cells, on the other hand, migrate through the ECM layer and cling to the bottom of the polycarbonate membrane. Invaded cells on the bottom of the insert membrane are dissociated from the membrane when incubated with Cell Detachment Buffer and subsequently lysed and detected by CyQuant GR dye.

      Application:

      The CHEMICON Cell Invasion Assay Kit is ideal for evaluation of invasive tumor cells. Each CHEMICON Cell Invasion Assay Kit contains sufficient reagents for the evaluation of 96 samples. The quantitative nature of this assay is especially useful for large scale screening of pharmacological agents.

      The CHEMICON Cell Invasion Assay Kit is intended for research use only; not for diagnostic or therapeutic 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.
      References
      Product Information
      Components
      • Sterile 96-well Cell Invasion Plate Assembly: (Part No. 90200) One 96-well feeder tray, and one 96-well Cell Invasion Chamber plate with ECMatrixTM -coated inserts.
      • 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 CHEMICON Cell Invasion Assay Kit employs a 96-well plate, and provides an efficient system for evaluating the invasion of tumor cells through a basement membrane model.
      Key Applications
      • Activity Assay
      Application NotesDetection of migrated cells is via

      CyQuant GR Dye®.
      Biological Information
      Species Reactivity
      • Human
      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 ConditionsStore kit materials at 2-8°C for up to their expiration date. Do not freeze.
      Packaging Information
      Material Size1 plate
      Material Package96 wells
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      QCM ECMatrix Cell Invasion Assay, 96-well (8 µm), fluorimetric SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Lentivirus-mediated RNA silencing of c-Met markedly suppresses peritoneal dissemination of gastric cancer in vitro and in vivo.
      Xiao-Lei Wang,Xi-Mei Chen,Jian-Ping Fang,Chang-Qin Yang
      Acta pharmacologica Sinica  33  2012

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      22407230 22407230
      Increases in mitochondrial biogenesis impair carcinogenesis at multiple levels.
      Xiao Wang,Carlos T Moraes
      Molecular oncology  5  2011

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      21855427 21855427
      Targeting of Integrin {beta}1 and Kinesin 2{alpha} by MicroRNA 183.
      Li G, Luna C, Qiu J, Epstein DL, Gonzalez P
      The Journal of biological chemistry  285  5461-71  2010

      Show Abstract Full Text Article
      19940135 19940135
      Differential expression of matrix metalloproteinases in brain- and bone-seeking clones of metastatic MDA-MB-231 breast cancer cells.
      Andreas M Stark,Bartosz Anuszkiewicz,Rolf Mentlein,Toshiyuki Yoneda,H Maximilian Mehdorn,Janka Held-Feindt
      Journal of neuro-oncology  81  2007

      Show Abstract
      16850107 16850107
      Genetic regulators of large-scale transcriptional signatures in cancer
      Adler, Adam S, et al
      Nat Genet, 38:421-30 (2006)  2006

      16518402 16518402
      New functions for the matrix metalloproteinases in cancer progression.
      Egeblad, Mikala and Werb, Zena
      Nat. Rev. Cancer, 2: 161-74 (2002)  2002

      Show Abstract
      11990853 11990853
      Sodium arsenite inhibits migration of extravillous trophoblast cells in vitro.
      Chi-Shan Li,Rita Loch-Caruso
      Reproductive toxicology (Elmsford, N.Y.)  24  2001

      Show Abstract
      17646079 17646079
      Transmembrane motility assay of transiently transfected cells by fluorescent cell counting and luciferase measurement.
      Gildea, J J, et al.
      BioTechniques, 29: 81-6 (2000)  2000

      Show Abstract
      10907081 10907081
      Tumor invasion and metastases: role of the basement membrane. Warner-Lambert Parke-Davis Award lecture.
      Liotta, L A
      Am. J. Pathol., 117: 339-48 (1984)  1984

      6095669 6095669

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      Categories

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