ECM508 Sigma-AldrichQCM Chemotaxis Cell Migration Assay, 24-well (8 µm), colorimetric
The QCM 24-well Migration Assay is ideal for the study of chemotaxis cell migration. The assay uses a 24-well plate with an 8 micron pore size, with colorimetric detection.More>> The QCM 24-well Migration Assay is ideal for the study of chemotaxis cell migration. The assay uses a 24-well plate with an 8 micron pore size, with colorimetric detection. Less<<
MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Key Specifications Table
|Key Applications||Detection Methods|
|Description||QCM Chemotaxis Cell Migration Assay, 24-well (8 µm), colorimetric|
|Overview||Also available: Cell Comb™ Scratch Assay! Get biochemical data from a scratch assay! Click Here
Cell migration is a fundamental function of normal cellular processes, including embryonic development, angiogenesis, wound healing, immune response, and inflammation. Microporous membrane inserts are widely used for cell migration and invasion assays. The most widely accepted of which is the Boyden Chamber assay. 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 migration. The effect of this treatment on cell behavior/migration remains questionable.
The CHEMICON® QCM™ 24-well Cell Migration Assay (ECM508) eliminates cell pre-labeling and manual counting. The 24-well insert and colorimetric detection format allows for quantitative comparison of multiple samples.
In the CHEMICON® QCM™ 24-well Migration Assay, cells that have migrated to the bottom of the insert membrane are stained. The stain is then extracted and transferred to a 96-well microtiter plate for colorimetric measurement.
The CHEMICON® QCM™ 24-well Migration Assay provides a quick and efficient system for quantitative determination of various factors on cell migration, including screening of pharmacological agents, evaluation of integrins or other adhesion receptors responsible for cell migration, or analysis of gene function in transfected cells.
The CHEMICON® QCM™ 24-well Migration Assay utilizes an 8 mm pore size, as this is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells; however, it is not appropriate for lymphocyte migration experiments. The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.
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
The CHEMICON® QCM™ 24-well Cell Migration Assay is performed in a Migration Chamber, based on the Boyden chamber principle. Each kit contains 24 inserts; each insert utilizes an 8 mm pore size polycarbonate membrane, as this is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells; however, it is not appropriate for lymphocyte migration experiments. Cells that have migrated through the polycarbonate membrane are incubated with Cell Stain Solution, then subsequently extracted and detected on a standard microplate reader (560 nm). The system may be adapted to study different types of cell migration, including haptotaxis, random migration, chemokinesis, and chemotaxis.
|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. Microplate reader (560 nm).
13. 24-well tissue culture plate.
14. Sterile cell culture hood.
|Safety Information according to GHS|
|Storage and Shipping Information|
|Storage Conditions||Store kit materials at 2-8°C for up to their expiration date. Do not freeze.|
|Material Size||1 plate|
|Material Package||24 wells|
QCM Chemotaxis Cell Migration Assay, 24-well (8 µm), colorimetric SDS
QCM Chemotaxis Cell Migration Assay, 24-well (8 µm), colorimetric Certificates of Analysis
|QCM 24-Well Colorimetric Cell Migration Assay - 3318231||3318231|
|Reference overview||Pub Med ID|
|The L6 domain tetraspanin Tm4sf4 regulates endocrine pancreas differentiation and directed cell migration.|
Keith R Anderson,Ruth A Singer,Dina A Balderes,Laura Hernandez-Lagunas,Christopher W Johnson,Kristin B Artinger,Lori Sussel
Development (Cambridge, England) 138 2011
The homeodomain transcription factor Nkx2.2 is essential for pancreatic development and islet cell type differentiation. We have identified Tm4sf4, an L6 domain tetraspanin family member, as a transcriptional target of Nkx2.2 that is greatly upregulated during pancreas development in Nkx2.2(-/-) mice. Tetraspanins and L6 domain proteins recruit other membrane receptors to form active signaling centers that coordinate processes such as cell adhesion, migration and differentiation. In this study, we determined that Tm4sf4 is localized to the ductal epithelial compartment and is prominent in the Ngn3(+) islet progenitor cells. We also established that pancreatic tm4sf4 expression and regulation by Nkx2.2 is conserved during zebrafish development. Loss-of-function studies in zebrafish revealed that tm4sf4 inhibits α and β cell specification, but is necessary for ε cell fates. Thus, Tm4sf4 functional output opposes that of Nkx2.2. Further investigation of how Tm4sf4 functions at the cellular level in vitro showed that Tm4sf4 inhibits Rho-activated cell migration and actin organization in a ROCK-independent fashion. We propose that the primary role of Nkx2.2 is to inhibit Tm4sf4 in endocrine progenitor cells, allowing for delamination, migration and/or appropriate cell fate decisions. Identification of a role for Tm4sf4 during endocrine differentiation provides insight into islet progenitor cell behaviors and potential targetable regenerative mechanisms.
|Plasma rich in growth factors (PRGF-Endoret) stimulates proliferation and migration of primary keratocytes and conjunctival fibroblasts and inhibits and reverts style=background-c|
Anitua E, Sanchez M, Merayo-Lloves J, De la Fuente M, Muruzabal F, Orive G
Investigative ophthalmology & visual science 52 6066-73. Print 2011 Aug. 2011
|Substance P Modulates Chronic Inflammation-Induced Colonic Fibrosis.|
Koon HW, Shih D, Karagiannides I, Zhao D, Fazelbhoy Z, Hing T, Xu H, Lu B, Gerard N, Pothoulakis C
Am J Pathol 2010
Substance P (SP) and the neurokinin-1 receptor (NK-1R) are involved in the development of colitis and mucosal healing after colonic inflammation. We studied whether SP modulates colonic fibrosis by using a chronic model of trinitrobenzenesulfonic acid (TNBS)-induced colitis in wild-type (WT) and NK-1R-deficient (NK-1R KD) mice. We found increased mRNA expression levels of collagen, vimentin, and the fibrogenic factors transforming growth factor β1 and insulin-like growth factor 1 in the chronically inflamed colons of WT mice treated with repeated intracolonic TNBS administrations. Fibrosis in TNBS-treated mice was also evident immunohistochemically by collagen deposition in the colon. Treatment of TNBS-exposed WT mice with the NK-1R antagonist CJ-12255 reduced colonic inflammation, colonic fibrosis, fibroblast accumulation, and expression levels of the fibrogenic factors. NK-1R knockout mice chronically exposed to TNBS had similar colonic inflammation compared with WT, but reduced colonic fibrosis, fibroblast accumulation, and expression levels of fibrogenic factors. Immunohistochemical staining also showed co-localization of NK-1R with fibroblasts in inflamed colons of mice and in colonic mucosa of patients with Crohn's disease. Exposure of human colonic CCD-18Co fibroblasts to SP (10 nmol/L) increased cell migration. SP stimulated collagen synthesis in CCD-18Co fibroblasts in the presence of transforming growth factor β1 and insulin-like growth factor 1, and this effect was reduced by Akt inhibition. Thus, SP, via NK-1R, promotes intestinal fibrogenesis after chronic colitis by stimulating fibrotic responses in fibroblasts.
|Presenilin 1/gamma-secretase is associated with cadmium-induced E-cadherin cleavage and COX-2 gene expression in T47D breast cancer cells.|
Chang Seok Park,Ohn Soon Kim,Sang-Moon Yun,Sangmee A Jo,Inho Jo,Young Ho Koh
Toxicological sciences : an official journal of the Society of Toxicology 106 2008
Cadmium is a heavy metal that has multiple toxic effects on human health and has been classified as a human carcinogen. E-cadherin is a major target of cadmium; however, the roles of E-cadherin and cadmium and the mechanisms of tumor progression remain to be defined. Here, we demonstrate that cadmium increases E-cadherin processing via a gamma-secretase in the T47D breast cancer cell lines. This presenilin 1 (PS1)/gamma-secretase-dependent cleavage of E-cadherin was accompanied by changes in reactive oxygen species or calcium. E-cadherin cleavage was blocked by a PS1 dominant-negative mutant, gamma-secretase inhibitors [N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester (DAPT) and L-685,486], antioxidants (N-acetylcysteine and Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin pentachloride), or a calcium chelating drug 1,2-bis(o-Aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester. Immunofluorescence analysis confirmed the disappearance of E-cadherin staining at the cell surface. Those inhibitors attenuated cadmium-induced cytotoxicity. Additionally, cadmium treatment increased cell motility and invasion ability, which was abated by DAPT. Interestingly, cyclooxygenase-2 (COX-2) expression induced by cadmium was also inhibited by DAPT. The cadmium-induced cell motility and invasion ability were inhibited by a COX-2 inhibitor, NS398. Our data indicate a novel molecular mechanism that links cytotoxicity of cadmium and disrupted E-cadherin processing to adherens junctions; cadmium induces COX-2 expression via gamma-secretase, which increases cell motility and invasion ability. Understanding the downstream signaling cascades of cadmium that promote tumor progression might be a key to the development of novel therapeutic strategies.