Sigma-Aldrich Milli ╡-Angiogenesis Activation Assay

Angiogenesis, the formation of new blood vessels from a pre-existing vascular network, occurs normally in development and is critical for a majority of the vessel formation that occurs during embryogenesis, tissue generation, and wound healing. However, abnormal blood vessel growth, either excessive or insufficient, can be the underlying cause of many deadly and debilitating diseases including cancer, cardiovascular disease, stroke, and diabetic and age-related blindness. Identification of specific compounds that promote or inhibit angiogenesis may provide promising treatments for these diseases.

Tubular formation is a multi-step process involving cell adhesion, migration, differentiation, and growth. Millipore′s TM=["Millicell"] µ-Angiogenesis Inhibition Assay provides an efficient system for the rapid and accurate identification of factors that inhibit tube formation of endothelial cells. The µ-Angiogenesis slide, with enhanced optical capabilities in a microscale format, allows for real time and continuous monitoring of tube formation in a simplifed and reproducible manner.

1. Millicell^® µ-Angiogenesis Slides (Part No. CS203030) should be stored at room temperature until ready to use.2. Fibrinogen Solution (Part No. CS203035) should be stored at -20°C and used within six months from the date of receipt. For short-term storage, store at room temperature for up to 2 weeks. Do not store at 4°C, as Fibrinogen Solutions tend to form aggregates when stored at 4°C. For long-term storage, aliquot into sterile tubes in a sterile laminar flow hood, freeze on dry ice, and store at -20°C.3. Thrombin Solution (Part No. CS203036) should be stored at -20°C and used within six months from the date of receipt. Unused Thrombin Solution can be stored at 4°C for up to 1 month. For longer term storage, Thrombin Solution may be stored in aliquots at -20°C.4. ITS Media Supplement, 100X (Part No. CS203037) should be stored at -20°C. Unused portions may also be stored at 4°C for up to 1 year. Avoid freeze thaw cycles.5. PMA (Phorbol-12-Myristate-13-Acetate) (Part No. CS203029) is provided as a 1 mg lyophilized powder. To prepare a 1 mg/mL stock, add 1 mL sterile DMSO (not provided). Vortex vigorously. Centrifuge briefly to collect the solution. Unused PMA Solution may be stored at -20°C in the dark for at least six months. WARNING: Highly irritating to skin and mucous membranes. TUMOR PROMOTER. Read the Material Safety Data Sheet (MSDS) prior to rehydration and use. Gloves and mask should be worn when using this compound.6. Calcein AM (Part No. CS202541) is provided as a 50 µg lyophilized powder. To prepare 1 mm Calcein AM Stock Solution, briefly centrifuge before opening the tube and then add 40 µL sterile DMSO (not provided). Vortex vigorously. Centrifuge briefly to collect the solution. Unused Calcein AM Solution may be stored in aliquots at –20°C for up to 1 month.

1. Millicell^® µ-Angiogenesis Slides: (Part No. CS203030) Five tissue culture treated slides. Each slide contains 15 wells. 2. Fibrinogen Solution: (Part No. CS203035) One 1.5 mL vial. 3. Thrombin Solution: (Part No. CS203036) One 0.75 mL vial. 4. ITS Media Supplement, 100X: (Part No. CS203037) One 0.5 mL vial. 5. PMA (Phorbol 12-Myristate 13-Acetate): (Part No. CS203029): One 1 mg vial. 6. Calcein-AM: (Part No. CS202541) One 50 µg vial.

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