Product Details
Cat # +Size | K457-100 |
---|---|
Size | 100 assays |
Kit Summary | • Fluorescence (Ex/Em: 365/410 nm) • Study and quantitate the effect of different compounds, proteins and tissue extracts on Actin polymerization and/or depolymerization • Evaluation of critical concentrations of actin polymerization in different conditions |
Detection Method | Fluorescence (Ex/Em: 365/410 nm) |
Species Reactivity | Not Applicable |
Applications | • Study and quantitate the effect of different compounds, proteins and tissue extracts on Actin polymerization and/or depolymerization • Evaluation of critical concentrations of actin polymerization in different conditions |
Kit Components | • Buffer G • Buffer P (10X) • Labeled Rabbit Muscle Actin • ATP (100 mM) |
Storage Conditions | -20°C |
Shipping Conditions | Gel Pack |
USAGE | For Research Use Only! Not For Use in Humans. |
Details
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1. Prepare an extraction buffer consisting of 20 mM Tris-HCl, pH 7.5, and 20 mM NaCl, plus any co-factors for your protein and a protease inhibitor cocktail.
2. Isolate extracts with ice-cold buffer and lyse the cells either using a 25 g bent over syringe needle or other device.
3. Make a 10 mg/ml protein extract with the ice-cold buffer.
4. Add 1/3rd volume of the extract to 2/3rd volume of pyrene-actin (labelled actin) to insure there is high enough concentration of protein to exert an effect on actin polymerization.
5. Prepare extract from experimental and control cells. The control cell line is very critical because the actin polymerization reaction is very sensitive to slight differences in protein concentration and salt.