ECM554 QCM ECMatrix Cell Invasion Assay, 24-well (8 µm), fluorimetric

ECM554
24 assays  24 assays
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      Overview

      Replacement Information

      Key Specifications Table

      Detection Methods
      Fluorescent
      Description
      Catalogue NumberECM554
      Brand Family Chemicon®
      Trade Name
      • QCM
      • Chemicon
      DescriptionQCM ECMatrix Cell Invasion Assay, 24-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 (1).

      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™ 24-well Invasion 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 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) (2-3). This green-fluorescent dye exhibits strong fluorescence enhancement when bound to cellular nucleic acids (4).

      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 (5-8). We examined the kit's performance using human fibrosarcoma (HT-1080) and non-invasive fibroblasts (NIH3T3).

      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 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.

      The ability to study cell invasion through an ECM barrier, is of vital importance for developing possible metastatic inhibitors and therapeutics. The new CHEMICON® QCM™ 24-well Invasion Assay (ECM554) provides an efficient, in vitro system for quantitative analysis of tumor cell invasion.

      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
      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 24-well Cell Invasion Plate Assembly: (Part No. 70019) Two 24-well plates with 12 ECMatrixTM-coated inserts per plate (24 coated inserts total/kit for 24 assays).
      • Cell Detachment Solution: (Part Number 90131) One bottle - 16 mL.
      • 4X Cell Lysis Buffer: (Part Number 90130) One bottle - 16 mL.
      • CyQuant GR Dye ®1: (Part Number 90132) One vial - 75 mL
      • Forceps: (Part Number 10203) One each.
      Detection methodFluorescent
      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 assays
      Material Package24 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      QCM ECMatrix Cell Invasion Assay, 24-well (8 µm), fluorimetric SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewSpeciesPub Med ID
      RNAi-mediated silencing of vEGF-C inhibits non-small cell lung cancer progression by simultaneously down-regulating the cXCR4, cCR7, vEGFR-2 and vEGFR-3-dependent axes-induced ERK, p38 and AKT signalling pathways.
      Feng Y, Hu J, Ma J, Feng K, Zhang X, Yang S, Wang W, Zhang J, Zhang Y
      European journal of cancer (Oxford, England : 1990)  2011

      Show Abstract
      21680174 21680174
      Modelling genetic and clinical heterogeneity in epithelial ovarian cancers.
      Lawrenson K, Sproul D, Grun B, Notaridou M, Benjamin E, Jacobs IJ, Dafou D, Sims AH, Gayther SA
      Carcinogenesis  2011

      Show Abstract
      21859834 21859834
      LIM and SH3 protein 1 (Lasp1) is a novel p53 transcriptional target involved in hepatocellular carcinoma.
      Bei Wang,Ping Feng,Ziwei Xiao,Ee-Chee Ren
      Journal of hepatology  50  2009

      Show Abstract
      19155088 19155088
      Screening and identification of novel B cell epitopes in human heparanase and their anti-invasion property for hepatocellular carcinoma.
      Jian-min Yang, Hui-ju Wang, Ling Du, Xiao-mei Han, Zai-yuan Ye, Yong Fang, Hou-quan Tao, Zhong-sheng Zhao, Yong-lie Zhou
      Cancer immunology, immunotherapy : CII  58  1387-96  2009

      Show Abstract
      19169879 19169879
      Gonadotropin-releasing hormone agonists suppress melanoma cell motility and invasiveness through the inhibition of alpha3 integrin and MMP-2 expression and activity.
      Moretti, Roberta M, et al.
      Int. J. Oncol., 33: 405-13 (2008)  2008

      Show Abstract
      Human18636163 18636163
      Cyr61/CCN1 is a tumor suppressor in human hepatocellular carcinoma and involved in DNA damage response.
      Ping Feng,Bei Wang,Ee Chee Ren
      The international journal of biochemistry & cell biology  40  2008

      Show Abstract
      17698398 17698398
      Genetic regulators of large-scale transcriptional signatures in cancer
      Adler, Adam S, et al
      Nat Genet, 38:421-30 (2006)  2006

      16518402 16518402

      Data Sheet

      Title
      QCM ECMatrix Cell Invasion Assay, Fluorimetric - Data Sheet

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