Actin-Toolkit Fluorometry with Pyrene Actin
α-Actin, Rabbit Skeletal Muscle
Cat. # 8030-01
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Applications
- Kinetic analysis of purified ligands, cell or tissue extracts on polymerization or depolymerization of actin;
- Determination of sensitivity to divalent cations or pharmacological compounds during polymerization or depolymerization;
- Measurement of F-actin severing effects of purified ligands, cell or tissue extracts at steady-state;
Description
|
Protein |
Pyrene Actin (α-Actin) |
|
Origin |
Skeletal muscle, rabbit |
|
Molecular mass |
42kDa |
|
Toolkit description |
The Actin-Toolkit Fluorometry is used to determine the polymerization and depolymerization reaction of actin by measuring the fluorescence of pyrenyl conjugated actin. By this means it also allows to analyze the interaction of actin with ligands or other components effecting the polymerization or depolymerization. The fluorescence of Pyrene Actin is maximally increased by a factor of 22 upon polymerization. The Actin-Toolkit Fluorometry is suitable to measure the effect on actin polymerization of purified ligands, cell or tissue extracts.
The Actin-Toolkit Fluorometry all-in-one kit contains components to measure the effects of native or recombinant proteins or fragments, lipids or other ligands on actin dynamics. The kit contains 4x1mg 10% pyrenyl actin labelled at Cys 374 with N-(1-pyrene) iodoacetamide. Buffers to keep actin functional and to vary e.g. the Ca++-concentration are also included for best results.
A handbook with a detailed description of experimental setups guides the user for safe, error-free handling of pyrenyl actin. We guarantee stable performance of the kit components for 6 months when stored at -70°C upon arrival. Experienced users may supplement this kit with 100% Pyrene Actin (dye/protein molar ratio = 1:1), to increase weak signals. |
|
Purity |
>99% by scanning densitometry. |
Properties
|
Form |
Lyophilized, ready-to-use. |
|
Kit content |
4x1.0mg Pyrene Actin 10% (lyophilized powder); |
|
Actin Buffer |
2mM Tris-Cl pH 8.2, 0.4mM ATP, 0.5mM DTT, 0.1mM CaCl2, 1mM NaN3 and 0.3% disaccharides, when reconstituted with 1.0 ml ultrapure water to obtain a 1mg/ml solution. |
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Related products |
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Purification notes |
Purified from rabbit skeletal muscle, GPC. |
|
Protein concentration |
Determined by the Biuret method. |
|
Storage instructions |
We guarantee stable performance of the kit components for 6 months when stored at -70°C upon arrival. Solubilized G-actin is kept on ice and should be used within 1 week. Avoid refreezing. |
|
Shipping conditions |
At ambient temperature. Upon delivery store at -70°C. |
|
Remarks |
For Use in Research only. Not for Use in Human or Veterinary Diagnostical or Therapeutical Applications. |
|
CAS no. |
51005-14-2 (Actin) |
Further Information
Material and Safety Data Sheet
Protein Sequence on NCBI
Handbook Actin-Toolkit Fluorometry
(Web version only for demonstration purpose!)
A full print version with protocols come with the kit.
Flyer Actin-Toolkits
Material and Safety Data Sheet
Protein Sequence on NCBI
