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ECM205

Sigma-Aldrich

Millicoat® ECM Screening Kit, 1 ea. ECM101-ECM105

Millicoat®, pkg of  96-well plate(s) (for fibronectin, vitronectin, laminin, collagen I & collagen IV)

Synonym(s):

Formerly under the CytoMatrix brand name.

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About This Item

UNSPSC Code:
12352202
eCl@ss:
32161000
NACRES:
NA.77

species reactivity

human

Quality Level

manufacturer/tradename

Chemicon®
Millicoat®

packaging

pkg of  96-well plate(s) (for fibronectin, vitronectin, laminin, collagen I & collagen IV)

technique(s)

activity assay: suitable
cell based assay: suitable

input

sample type neural stem cell(s)
sample type: human embryonic stem cell(s)
sample type: mouse embryonic stem cell(s)
sample type epithelial cells
sample type hematopoietic stem cell(s)
sample type pancreatic stem cell(s)
sample type induced pluripotent stem cell(s)
sample type mesenchymal stem cell(s)

detection method

colorimetric

shipped in

wet ice

General description

Cell adhesion plays a major role in cellular communication and regulation, and is of fundamental importance in the development and maintenance of tissues. Scientists are continually examining the adhesion and migration of many diverse cell types on various extracellular matrix (ECM) component proteins. Millicoat® Cell Adhesion Strips are provided as 12 8-well removable strips in a plate frame for convenience and flexibility in designing assays. The wells in rows A - G have been coated with a human ECM protein. Row H of each strip is coated with BSA which serves as a negative assay control. The Millicoat screening kit contains individual 96-well plates for fibronectin, vitronectin, laminin, collagen I and collagen IV. Cells are seeded onto the coated substrate. Subsequently, adherent cells are fixed and stained. Relative attachment is determined using absorbance readings.

Application

PROCEDURE:

NOTE: Optimal assay performance is obtained using subconfluent cell cultures. This can be achieved by splitting the cells 1 to 2 days prior to performing the assay.

1. Rehydrate the strips with 200 mL of PBS per well for at least 15 minutes at room temperature. Remove the PBS from the rehydated strips.

2. Prepare a single cell suspension, preferably using a non-enzymatic dissociation buffer. Optimum cell density may be determined by titration of the cells. A common starting range is between 1x10E05 to 1x10E07 cells/mL.

3. Add 100 mL of the diluted cell suspension to each well. Incubate the plate at 37°C for 1 hour in a CO2 incubator. Gently wash the plate 3 times with PBS containing Ca2+/Mg2+ (200 mL/well).

4. Add 100 mL/well of 0.2% crystal violet in 10% ethanol to each well. Incubate for 5 minutes at room temperature. Remove the stain from the wells. Gently wash the plate 3 times with PBS (300 mL/well).

5. Add 100 mL of Solubilization Buffer (A 50/50 mixture of 0.1M NaH2PO, pH 4.5 and 50% ethanol) to each well. Allow strips to incubate and gently shake at room temperature until the cell-bound stain is completely solubilized; approximately 5 minutes.

6. Determine the absorbances at 540 - 570 nm on a microplate reader.
Research Category
Cell Structure
The Millicoat® screening kit contains individual 96-well plates for fibronectin, vitronectin, laminin, collagen I & collagen IV.

Storage and Stability

May be stored at 2-8°C in the foil pouch for at least 3 months. Unused strips may be placed back in the pouch for storage. Ensure that the desiccant remains in the pouch, and that the pouch is securely closed.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
MILLICOAT is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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 Class Code

11 - Combustible Solids


Certificates of Analysis (COA)

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Mary Ann Stepp et al.
Journal of cell science, 120(Pt 16), 2851-2863 (2007-08-02)
We have reported previously that syndecan-1 (Sdc1)-null mice show delayed re-epithelialization after skin and corneal wounding. Here, we show that primary keratinocytes obtained from Sdc1-null mice and grown for 3-5 days in culture are more proliferative, more adherent and migrate
K E Crawford et al.
Placenta, 33(5), 416-423 (2012-03-02)
Calreticulin is a calcium binding, endoplasmic reticulum resident protein best known for its roles in intracellular calcium homeostasis and the quality control processes of the endoplasmic reticulum. There is evidence for a range of activities for calreticulin outside the endoplasmic
Lichun Sun et al.
Journal of Cancer, 7(11), 1388-1395 (2016-07-30)
Cervical cancer is a second leading cancer death in women world-wide, with most cases in less developed countries. Notch signaling is highly conserved with its involvement in many cancers. In the present study, we established stable cervical cell lines with
Yuanyuan Hua et al.
Oncotarget, 7(40), 66077-66086 (2016-09-08)
Epithelial ovarian carcinoma accounts for 90% of all ovarian cancer and is the most deadly gynecologic malignancy. Recent studies have suggested that fallopian tube fimbriae can be the origin of cells for high-grade serous subtype of epithelial ovarian carcinoma (HGSOC).
Stephanie Arndt et al.
The American journal of pathology, 178(6), 2622-2631 (2011-06-07)
Dermal wound healing depends on highly complex interplay among various cytokines and cell types. Disruption of this process can result in impaired healing in the form of excessive scarring, as is the case in fibrotic diseases such as keloid and

Protocols

This page covers the ECM coating protocols developed for four types of ECMs on Millicell®-CM inserts, Collagen Type 1, Fibronectin, Laminin, and Matrigel.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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