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ECM551 | QCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric

ECM551
24 wells  24 wells
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      Panorama geral

      Replacement Information

      Tabela com principais espec.

      Detection Methods
      Chromogenic
      Description
      Catalogue NumberECM551
      Brand Family Chemicon®
      Trade Name
      • QCM
      • Chemicon
      DescriptionQCM™ Collagen Cell Invasion Assay, 24-well (8 µm), Colorimetric
      OverviewNEW!! High Sensitivity Collagen Invasion Assay! 24-well (8 µm), Colorimetric Click Here

      Also 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. 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™ 24-well Collagen-based Invasion Assay (ECM551) provides an efficient, in vitro system for quantitative analysis of tumor cell invasion.

      The CHEMICON QCM™ 24-well Collagen-based Invasion Assay (ECM551) eliminates cell pre-labeling and manual counting. The 24-well insert and colorimetric detection format allows for quantitative comparison of multiple samples. ECM551 uses purified chicken type I collagen as a matrix. The product is 99.9% pure native atelomeric avian collagen. Approximately 85% type I and 15% type III collagen protein. The material has been specially prepared to maintain its native state as much as possible. The telomeric ends of the collagen I have been enzymatically removed. Since most of the species identity is found within the telomeric ends, our atelomeric chicken collagen should provide a suitable substrate for most any animal species. The working concentration and precise purification protocol are proprietary and will not be disclosed, however it at least a 0.3% solution of treated avian collagen.

      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 Cell Invasion Assay is performed in an Invasion Chamber, based on the Boyden chamber principle. Each kit contains 24 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 incubated with Cell Stain Solution, then subsequently extracted and detected on a standard microplate reader (560 nm).
      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. Microplate reader (560 nm).

      13. 24-well tissue culture plate.

      14. Sterile cell culture hood.
      References
      Product Information
      Components
      • Sterile 24-well Cell Invasion Plate Assembly: (Part No. 90248) Two 24-well plates with 12 collagen-coated inserts per plate (24 inserts total/kit).
      • Cell Stain: (Part No. 90144) One 20 mL bottle.
      • Extraction Buffer: (Part No. 90145) One 20 mL bottle.
      • Cotton Swabs: (Part No. 10202) 50 each.
      • Forceps: (Part No. 10203) One each.
      Detection methodChromogenic
      Applications
      ApplicationThe Cell Invasion Assay Kit kuses a 24 well plate with 8 um pores, which 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 24 samples. The quantitative nature of this assay is especially useful for 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 Size24 wells
      Material Package24 wells
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Título

      Ficha de Segurança de Produtos (MSDS) 

      Referências

      Visão geral das referênciasPub Med ID
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      Cell cycle (Georgetown, Tex.) 11 2012

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      Molecular cancer 11 2012

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      Plumbagin, a plant derived natural agent inhibits the growth of pancreatic cancer cells in in vitro and in vivo via targeting EGFR, Stat3 and NF-κB signaling pathways.
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      International journal of cancer. Journal international du cancer 131 2012

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      TRAIL-induced caspase/p38 activation is responsible for the increased catalytic and invasive activities of Akt.
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      International journal of oncology 38 2011

      Mostrar Resumo Texto completo do artigo
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      Inhibition of adhesion, invasion, and metastasis by antibodies targeting CEACAM6 (NCA-90) and CEACAM5 (Carcinoembryonic Antigen).
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      Nat. Rev. Cancer, 2: 161-74 (2002) 2002

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      A new in vitro assay for quantitating tumor cell invasion.
      Repesh, L A
      Invasion Metastasis, 9: 192-208 (1989) 1989

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      A rapid in vitro assay for quantitating the invasive potential of tumor cells.
      Albini, A, et al.
      Cancer Res., 47: 3239-45 (1987) 1987

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      Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells.
      Terranova, V P, et al.
      Proc. Natl. Acad. Sci. U.S.A., 83: 465-9 (1986) 1986

      Mostrar Resumo
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      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

      Brochura

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

      Ficha de dados

      Título
      QCM Collagen Cell Invasion Assay, Colorimetric - Data Sheet
      QCM Collagen Cell Invasion Assay, Colorimetric - Data Sheet

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      Categorias

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