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- ECM554
- 2025年07月13日
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- Vertebrates
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- 宿主：
0
- 适应物种：
Vertebrates
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- 库存：
大量
- 是否单克隆：
0
- 规格：
1 plate
Description:
QCM ECMatrix Cell Invasion Assay, 24-well (8 μm), fluorimetric
Trade Name:
- QCM
- Chemicon (Millipore)
Product Overview:
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
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
Application Notes:
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 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.
The CHEMICON® Cell Invasion Assay Kit is intended for research use only; not for diagnostic or therapeutic applications.
Species Reactivity:
Vertebrates
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.
Kit or Assay Type:
Cell Invasion Assays
Components:
- Sterile 24-well Cell Invasion Plate Assembly: (Part No. 70019) Two 24-well plates with 12 ECMatrixTM-coated inserts per plate (24 inserts total/kit).
- 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.
Storage Conditions:
Store kit materials at 2-8°C for up to their expiration date. Do not freeze.
Detection Methods:
Fluorescent
Packaging:
24 wells
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.
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.
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详细描述见链接：http://www.millipore.com/catalogue/item/ECM554
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QCM ECMatrix Cell Invasion Assay, Fluorimetric; 1 plate
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