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ECM556 QCM Collagen Cell Invasion Assay, 96-well (8 µm), fluorimetric

The Cell Invasion Assay Kit is ideal for evaluation of invasive tumor cells.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
ECM556
1 plate  96 wells
Purchase on Sigma-Aldrich

概述

Replacement Information

重要规格表

Detection Methods
Fluorescent
Description
Catalogue NumberECM556
Brand Family Chemicon®
Trade Name
  • QCM
  • Chemicon
DescriptionQCM Collagen Cell Invasion Assay, 96-well (8 µm), fluorimetric
OverviewAlso available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here

Introduction
Penetration of the subendothelial basement membrane marks a critical turning point in the metastatic process. As proliferating neoplastic cells attempt to escape the primary tumor site, local invasion of the surrounding tissue (interstitial stroma) must occur. Neovascularization is initiated by expression of angiogenic factors (e.g. FGF, VEGF, HGF), providing nutritional requirements and access to the vascular system. Prior to penetrating the blood vessel endothelium and gaining access to the blood stream (intravasation), cancer cells must invade local tissues by degrading ECM components and ultimately, transverse the basement membrane. Once in circulation, these cells can form metastatic colonies at secondary locations, making this membrane a key invasive barrier.

The basement membrane surrounding the blood vessel endothelium is a thin, specialized network of extracellular matrix proteins (ECM) that serves many functions. Comprised of proteins and proteoglycans, such as collagen, laminin, entactin, fibronectin, heparin sulfate and perlecan, this membrane acts as a physical barrier between the epithelium and underlying tissues. It provides cell surface anchorage (via integrins, receptor kinases, and cell surface proteoglycans), induces cellular differentiation, gives architectural support, and limits the migration of normal cells. The ability of tumor cells to degrade the ECM components of the basement membrane and surrounding tissues is directly correlated with metastatic potential. By releasing proteolytic enzymes (e.g. MMP collagenases, plasminogen activators, cathepsins), cancer cells are able to breach the membrane and penetrate the blood vessel wall (1). Collagen, the primary structural element of the basement membrane and tissue scaffolding protein, represents the main deterrent in the migration of tumor cells.

The ability to study cell invasion through a collagen barrier, is of vital importance for developing possible metastatic inhibitors and therapeutics. The new CHEMICON QCM™ 96-well Collagen-based Invasion Assay (ECM556) provides an efficient, in vitro system for quantitative, high-throughput analysis of tumor cell invasion.

In the QCM™ 96-well Collagen-based Invasion Assay (ECM556), invaded cells on the bottom of the insert membrane are dissociated from the membrane when incubated with a specially formulated Cell Detachment Buffer. The invasive cells are subsequently lysed and detected by the patented CyQuant dye (Molecular Probes) (2-3). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (4).



The CHEMICON QCM™ 96-well Collagen-based Invasion Assay (ECM556) eliminates cell pre-labeling, fixing/staining, swabbing, and manual counting. The 96-well insert and homogenous fluorescence detection format allows for high-throughput, quantitative comparison of multiple samples.

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)
  • 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 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 polymerized collagen. The collagen layer occludes the membrane pores, blocking non-invasive cells from migrating through. Invasive cells, on the other hand, migrate through the polymerized collagen 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.
    		•
    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. 90249) One 96-well feeder tray, and one 96-well Cell Invasion Chamber plate with collagen-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®1: (Part No. 90132) One vial - 75 mL
    Detection methodFluorescent
    HS Code3822 19 90
    Quality LevelMQ100
    Applications
    ApplicationThe Cell Invasion Assay Kit is ideal for evaluation of invasive tumor cells.
    Application NotesThe 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.
    Biological Information
    Species Reactivity
    • Vertebrates
    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
    Global Trade Item Number
    产品目录编号 GTIN
    ECM556 04053252335310

    Documentation

    QCM Collagen Cell Invasion Assay, 96-well (8 µm), fluorimetric MSDS

    职位

    物料安全数据表 (MSDS) 

    参考

    参考概述公共医疗ID
    A conjugate of camptothecin and a somatostatin analog against prostate cancer cell invasion via a possible signaling pathway involving PI3K/Akt, alphaVbeta3/alphaVbeta5 and MMP-2/-9.
    Li-Chun Sun, Jing Luo, L Vienna Mackey, Joseph A Fuselier, David H Coy
    Cancer letters  246  157-66  2007

    显示摘要     		16644105 16644105
    Effects of camptothecin on tumor cell proliferation and angiogenesis when coupled to a bombesin analog used as a targeted delivery vector.
    Li-Chun Sun,Jing Luo,Vienna L Mackey,Joseph A Fuselier,David H Coy
    Anti-cancer drugs  18  2007

    显示摘要     		17264768 17264768

    小册子

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

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