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Millicell® μ-Angiogenesis Activation Assay Kit
Description:
Millicell® μ-Angiogenesis Activation Assay Kit
Trade Name:
Millicell
Qty/Pk:
1 Kit
Product Overview:
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 Millicell® µ-Angiogenesis Activation Assay provides an efficient system for the rapid and accurate identification of factors that stimulate tube formation of endothelial cells. The Millicell µ-Angiogenesis Activation Assay takes advantage of the propensity of endothelial cells to rapidly align and form interconnecting networks that display patent lumina when cultured on top or within a fibrin gel. 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.
Tubular formation is a multi-step process involving cell adhesion, migration, differentiation, and growth. Millipore’s Millicell® µ-Angiogenesis Activation Assay provides an efficient system for the rapid and accurate identification of factors that stimulate tube formation of endothelial cells. The Millicell µ-Angiogenesis Activation Assay takes advantage of the propensity of endothelial cells to rapidly align and form interconnecting networks that display patent lumina when cultured on top or within a fibrin gel. 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.
Key Applications:
- Activity Assay
- Cell Culture
Species Reactivity:
- Vertebrates
- Rat
- Human
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.
Components:
- 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.
Storage Conditions:
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 -20ºC 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 4ºC, as Fibrinogen Solutions tend to form aggregates when stored at 4ºC. For long-term storage, aliquot into sterile tubes in a sterile laminar flow hood, freeze on dry ice, and store at -20ºC.
3. Thrombin Solution (Part No. CS203036) should be stored at -20ºC and used within six months from the date of receipt. Unused Thrombin Solution can be stored at 4ºC for up to 1 month. For longer term storage, Thrombin Solution may be stored in aliquots at -20ºC.
4. ITS Media Supplement, 100X (Part No. CS203037) should be stored at -20ºC. Unused portions may also be stored at 4ºC 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 -20ºC 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 –20ºC for up to 1 month.
2. Fibrinogen Solution (Part No. CS203035) should be stored at -20ºC 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 4ºC, as Fibrinogen Solutions tend to form aggregates when stored at 4ºC. For long-term storage, aliquot into sterile tubes in a sterile laminar flow hood, freeze on dry ice, and store at -20ºC.
3. Thrombin Solution (Part No. CS203036) should be stored at -20ºC and used within six months from the date of receipt. Unused Thrombin Solution can be stored at 4ºC for up to 1 month. For longer term storage, Thrombin Solution may be stored in aliquots at -20ºC.
4. ITS Media Supplement, 100X (Part No. CS203037) should be stored at -20ºC. Unused portions may also be stored at 4ºC 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 -20ºC 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 –20ºC for up to 1 month.
Detection Methods:
Fluorescent
Product Name:
Millicell® μ-Angiogenesis Activation Assay Kit
Materials Required but Not Delivered:
1. Inverted Light Microscope
2. Fluorescent microscope (if Calcein AM is used)
3. Precision pipettes
4. Accutase™ Cell Dissociation Solution (Catalog. No. SCR005)
5. Human umbilical vein endothelial cells (HUVEC) (Catalog No. SCCE001) or any other experimental cell lines capable of tube formation.
6. EndoGRO™ LS Complete Medium (Cat. No. SCME001) or other endothelial cell basal medium.
7. Phosphate-Buffered Saline (1X PBS) (Catalog No. BSS-1005-B)
8. Sterile distilled water (Cat. No. TMS-006-C or TMS-006-B)
9. Low speed centrifuge
10. Sterile eppendorf tubes
11. Precision pipette tips. We recommend 0.1–10 mL TipOne filter tips (USA Scientific Catalog No. 1121-3810) or Rainin’s 20 µL Precision Pipette Tips (Rainin Catalog No. RT-L10S)
12. 37ºC Incubator with 5% CO2
13. Hemacytometer
14. Trypan Blue
15. DMSO
2. Fluorescent microscope (if Calcein AM is used)
3. Precision pipettes
4. Accutase™ Cell Dissociation Solution (Catalog. No. SCR005)
5. Human umbilical vein endothelial cells (HUVEC) (Catalog No. SCCE001) or any other experimental cell lines capable of tube formation.
6. EndoGRO™ LS Complete Medium (Cat. No. SCME001) or other endothelial cell basal medium.
7. Phosphate-Buffered Saline (1X PBS) (Catalog No. BSS-1005-B)
8. Sterile distilled water (Cat. No. TMS-006-C or TMS-006-B)
9. Low speed centrifuge
10. Sterile eppendorf tubes
11. Precision pipette tips. We recommend 0.1–10 mL TipOne filter tips (USA Scientific Catalog No. 1121-3810) or Rainin’s 20 µL Precision Pipette Tips (Rainin Catalog No. RT-L10S)
12. 37ºC Incubator with 5% CO2
13. Hemacytometer
14. Trypan Blue
15. DMSO