Tropoelastin is a 60-72 kDa alternatively spliced extracellular matrix protein. It is a soluble precursor of elastin and is distributed in elastic tissues. Tropoelastin consists of an N- terminal 26-amino-acid signal peptide and highly basic strongly conserved C-terminal domain, that is required for both incorporation into elastin and for cellular interactions. It Is also characterized with a hydrophobic domain and exon 26A, which is an unusual highly hydrophilic domain.
Use this procedure as a guideline to determine optimal coating conditions for the culture system of choice.
1. Add 1 ml of sterile 0.25% glacial acetic acid solution to the tropoelastin serum vial containing 1 mg.
2. Mix contents gently until material is completely solubilized. The solution may remain slightly hazy.
3. Transfer desired volume of solution from the serum vial to a dilution vessel if required. Further dilute to desired concentration using sterile 0.25% Acetic Acid solution. A typical working concentration may range from 1 to 50 μg/ml.
4. Add appropriate amount of diluted tropoelastin material to the culture surface.
5. Incubate at room temperature or 37 °C, covered, for approximately 1.5 hours.
6. After incubation, aspirate any remaining material.
7. Rinse coated surfaces carefully with sterile water and avoid scratching surfaces.
8. Coated surfaces are ready for use. They may also be stored at 2-10 °C damp or air dried if sterility is maintained.
Note: To maintain sterility, perform all operations in a laminar flow hood employing aseptic techniques.
Tropoelastin exhibits the property of coacervation and plays a vital role in fiber formation and assembly. Abnormality in the tropoelastin is associated with the development of hereditary disease, cutis laxa (where the elasticity of the skin is lost).
Features and Benefits
Tropoelastin available in its native state. Unique since it becomes cross-linked immediately after its synthesis by the cell and during its export into the extracellular matrix.
Identity and purity qualitatively determined by SDS-PAGE
Storage at -20 °C prior to solubilization is recommended. Storage at 2-10 °C after reconstitution is recommended.
Shelf life is 24 months when stored at -20 °C. Shelf life after reconstitution is 3 months when stored at 2-10 °C