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ECM104 EMD Millipore

Millicoat™ Human Collagen Type I Coated Strips (96-Wells)

96-well plate coated with human Collagen Type I used for cell adhesion studies.

Synonym: Formerly under the CytoMatrixTM brand name.

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Properties

trade name   Millicoat
  Chemicon
brand family   Chemicon
detection method   Chromogenic
application(s)   Enzyme Assays
stem cell type   Human Embryonic Stem Cells
  Mouse Embryonic Stem Cells
  Mesenchymal Stem Cells
  Neural Stem Cells
  Hematopoietic Stem Cells
  Epithelial Cells
  Pancreatic Stem Cells
  Induced Pluripotent Stem Cells
species reactivity   Human
NCBI accession no.   NM_000088.3
UniProt accession no.   P02452
gene symbol   COL1A1(1277)
  OI4
material size   1 plate
shipped in   wet ice
storage conditions   Strips 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.

Description

Other Notes

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 x 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 Human Collagen Type I. Row H of each strip is coated with BSA which serves as a negative assay control. 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 timing and performance may vary for different cell lines but generally can be obtained using subconfluent cell cultures in the assay described below. Subconfluent cultures can be achieved by splitting 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 rehydrated 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 45 minutes in a CO2 incubator. Gently wash the plate 2-3 times with PBS containing Ca2+/Mg2+ (200 mL/well).

4. Add 100 mL 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 strips 3-5 times with PBS (300 mL/well) to remove the excess stain.

5. Add 100 mL of Solubilization Buffer (A 50/50 mixture of 0.1M NaH2PO4, 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 absorbance at 540 - 570 nm on a microplate reader.

material package

96 wells

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.

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Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis

Protocols & Articles

Articles

Extracellular Matrix Proteins and tools for cell culture optimization

Animal cells and tissue culture techniques are constantly improved to optimize in vitro cell culture conditions. Extracellular Matrix (ECM) proteins coating, chemical or physical modification of the ...
Keywords: Adhesion, Angiogenesis, Apoptosis, Asymmetric synthesis, Cancer, Cell attachment, Cell culture, Cell proliferation, Coagulation, Endocrinology, Growth factors, Hormones

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