CytoSelect™ 48-well Cell Adhesion Assay (ECM Array, Fluorometric)

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Applikation
Cellular Assay (CA)
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Marke CytoSelect™
Proben Serum, Cell Samples
Analytische Methode Quantitative
Nachweismethode Fluorometric
Produktmerkmale The CytoSelect™ Cell Adhesion Assay Kit provides a rapid, quantitative method for evaluating cell adhesion. The kit contains sufficient reagents for the evaluation of 48 samples (40 ECM protein-coated wells, 8 BSA-coated wells).
Bestandteile
  1. ECM Adhesion Plate : One 48-well plate containing 40 ECM protein-coated wells and 8 BSA-coated wells (see layout below). FN, Collagen IV and Fibrinogen are from human, Laminin I is from Mouse and Collagen I is from Bovine.
  2. 4X Lysis Buffer : One Bottle - 10.0 mL
  3. CyQuant® GR Dye : One tube - 50 μL 2
Benötigtes Material
  1. Cell culture medium
  2. Serum free medium, such as DMEM containing 0.5 % BSA, 2 mM CaCl2 and 2 mM MgCl2
  3. Cell culture incubator (37 °C, 5 % CO2 atmosphere)
  4. 1X PBS containing 2 mM CaCl2 and 2 mM MgCl2
  5. Light microscope
  6. 96-well plate suitable for a fluorescence plate reader
  7. Fluorescence plate reader
Hintergrund Cell adhesion is a complex process involved in embryogenesis, migration/invasion, tissue remodeling, and wound healing. To perform these processes, cells adhere to extracellular matrix components (via adhesion receptors), forming complexes with components of the cytoskeleton that ultimately affect cell motility, differentiation, proliferation, and survival.
Applikationshinweise Optimal working dilution should be determined by the investigator.
Kommentare

  • Full quantitation of cell adhesion with no manual cell counting
  • Plates precoated with a uniform substrate layer of a single ECM protein in each row: Collagen I, Collagen IV, Fibrinogen, Fibronectin, and Laminin

Plattentyp Pre-coated
Protokoll The CytoSelect™ Cell Adhesion Assay Kit utilizes an ECM protein-coated 48-well plate (see Adhesion Plate Layout). First, cells are seeded onto the coated substrate, where the adherent cells are captured. Next, unbound cells are removed with consecutive washes. Finally, the adherent cells are lysed and subsequently detected with CyQuant® GR Dye.
Testdurchführung
  1. Under sterile conditions, allow the ECM Adhesion Plate to warm up at room temperature for 10 minutes.
  2. Prepare a cell suspension containing 0.1-1.0 x 106 cells/mL in serum free media. Agents that inhibit or stimulate cell adhesion can be added directly to the cell suspension. 3
  3. Add 150 μL of the cell suspension to the inside of each well (BSA-coated wells are provided as a negative control).
  4. Incubate for 30-90 min in a cell culture incubator.
  5. Carefully discard or aspirate the media from each well (Note: Do not allow wells to dry). Gently wash each well 4-5 times with 250 μL PBS.
  6. Prepare sufficient 1X Lysis Buffer/CyQuant® GR dye solution for all samples by diluting the dye 1:300 in Lysis Buffer (for example, add 4 μL dye to 300 μL of 4X Lysis Buffer and 900 μL of dH2O).
  7. Add 200 μL of 1X Lysis Buffer/CyQuant® GR dye solution to each well containing cells. Incubate 20 minutes at room temperature with shaking.
  8. Transfer 150 μL of the mixture to a 96-well plate suitable for fluorescence measurement. Read fluorescence with a fluorescence plate reader at 480 nm/520 nm.
Beschränkungen Nur für Forschungszwecke einsetzbar
Lagerung 4 °C
Informationen zur Lagerung Store all kit components at 4°C.
Produkt verwendet in: Zohra, Medved, Lazareva, Polyak: "Functional behavior and gene expression of magnetic nanoparticle-loaded primary endothelial cells for targeting vascular stents." in: Nanomedicine (London, England), Vol. 10, Issue 9, pp. 1391-406, 2015 (PubMed).

Kim, Kim, Huang, Okada, Welge, Wang, Ashraf: "Cardiac stem cells with electrical stimulation improve ischaemic heart function through regulation of connective tissue growth factor and miR-378." in: Cardiovascular research, Vol. 100, Issue 2, pp. 241-51, 2013 (PubMed).