Actin-Toolkit Fluorometry Actin488
Fluorescent native α-actin from rabbit skeletal muscle
Cat. #: 8033-01
Applications
- Kinetic analysis of effects of purified ligands, cell extracts or tissue extracts on actin polymerization and depolymerization.
- Analysis of the influence of divalent cations or pharmacological compounds on actin assembly and disassembly.
- Measurement of F-actin severing or depolymerizing activity under steady-state conditions.
Description
| Product name | Actin-Toolkit Fluorometry Actin488 |
| Protein | Actin488 (α-actin) |
| Origin | Rabbit skeletal muscle |
| Molecular mass | 42 kDa |
| Toolkit description |
The Actin-Toolkit Fluorometry Actin488 is designed for fluorescence-based analysis of actin polymerization and depolymerization. Changes in Actin488 fluorescence can be followed during actin assembly and disassembly, providing a kinetic readout of actin dynamics under defined in vitro conditions. The assay can be used to examine the effects of purified proteins, recombinant fragments, lipids, pharmacological compounds, cell extracts or tissue extracts on actin polymerization and depolymerization. The kit contains 4 × 1.0 mg fluorescent rabbit skeletal muscle Actin488 together with Gelsolin, ATP-actin buffer components, PolyMix and CaCl2 for controlled variation of polymerization conditions. A dedicated handbook describes fluorometric actin-polymerization and depolymerization assays. |
| Optical properties | Actin488: excitation 495 nm, emission 520 nm |
| Purity | >99% by scanning densitometry. |
Properties
| Form | Lyophilized. |
| Kit content | 4 × 1.0 mg Actin488 (lyophilized powder); 1 × 20 µg Gelsolin; 1 × 1000 ml ATP-Actin Buffer, supplied as components; 1 × 1.0 ml PolyMix (10× stock); 1 × 0.5 ml CaCl2 (100 mM); 1 × Toolkit Handbook Fluorometry; 10 × black tubes and tags. |
| Actin buffer after reconstitution | 2 mM Tris-Cl, pH 8.2, 0.4 mM ATP, 0.5 mM DTT, 0.1 mM CaCl2 and 0.3% disaccharides when reconstituted with 1.0 ml ultrapure water to obtain a 1 mg/ml actin solution. |
| Related products | Actin-binding proteins and additional fluorescent actin reagents for defined in vitro assays. |
| Purification notes | Native actin purified from rabbit skeletal muscle; final purification by gel permeation chromatography (GPC). Actin488 is prepared from purified actin. |
| Protein concentration | Actin concentration is determined by direct UV absorbance at A290 nm with correction for fluorophore absorbance where applicable. For unlabeled actin, A290nm 1% = 0.62. |
| Storage instructions | We guarantee stable performance of the kit components for 6 months when stored at −70 °C upon arrival. Solubilized G-actin should be kept on ice and used within 1 week. Avoid refreezing. |
| Shipping conditions | Shipped at ambient temperature. Upon delivery store at −70 °C. |
| CAS no. | 51005-14-2 (Actin) |
| Remarks | For research use only. Not for diagnostic, therapeutic, clinical or in vivo use. |
Buffers and proteins from Hypermol® are made using ultrapure reagents in Milli-Q™ water, as described in our publications.
Further Information
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