Actin-Toolkit TIRFM
The complete solution!
ATTO488-labeled native α-actin from rabbit skeletal muscle
Cat. #: 8093-01
Applications
- Single-molecule analysis of actin polymerization, depolymerization and nucleation by TIRF microscopy.
- Visualization of ligand-induced changes in actin filament dynamics.
- Fluorescence-based analysis of actin filaments, bundles and networks under defined in vitro conditions.
Description
| Product name | Actin-Toolkit TIRFM | ATTO488-Actin |
| Protein | ATTO488-Actin (α-actin) |
| Origin | Rabbit skeletal muscle |
| Molecular mass | 42 kDa |
| Toolkit description |
The Actin-Toolkit TIRFM is designed for direct visualization of actin dynamics by total internal reflection fluorescence microscopy and related fluorescence-based single-molecule imaging approaches. The toolkit contains ATTO488-labeled rabbit skeletal muscle α-actin with a Degree of Labeling (DOL) of 0.3. The labeled actin is intended for incorporation into actin filaments at a low labeling fraction suitable for fluorescence imaging of filament assembly and dynamics. NEM-Myosin II is supplied as a surface-tethering component for flow-chamber assays. The kit also contains passivation solution, actin polymerization and monomer buffers, Mg-exchange buffer and FluMaXx oxygen-scavenging reagents for fluorescence imaging under reduced photochemical stress. A dedicated handbook describes flow-chamber assembly and TIRFM actin-polymerization assays. |
| Degree of Labeling | DOL: 0.3 |
| Optical properties | ATTO488: λabs 501 nm, λfl 523 nm |
| Purity | >99% by scanning densitometry. |
Properties
| Form | Lyophilized and in solution, depending on kit component. |
| Kit content | 5 × 100 µg ATTO488-Actin (lyophilized powder); 1 × 100 µg NEM-Myosin II (lyophilized powder); 3 × 1 ml Passivation solution (5% BSA, fatty-acid free); 1 × 1 ml PolyMix (lyophilized F-actin ATP buffer); 1 × 50 ml MonoMix (lyophilized G-actin ATP buffer); 1 × 2 ml HSB, 5× stock (Myosin Buffer, pH 7.0); 1 × 0.5 ml ME-Buffer (100× Mg-exchange buffer); 1 × Toolkit Handbook TIRFM. |
| FluMaXx components |
5 × 250 µl FluMaXx (2× stock, lyophilized);
Contains glucose oxidase, catalase, imidazole pH 7.4, KCl, EGTA, MgCl2, glucose and cryoprotectants (proprietary formula); 1 × 500 µl ATP/DTT Mix (10× stock, lyophilized); 2 × 1.5 ml methylcellulose stock solution (0.5% in Milli-Q™ water). |
| 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 actin reagents for defined in vitro reconstitution experiments. |
| Purification notes | Native actin purified from rabbit skeletal muscle; final purification by gel permeation chromatography (GPC). ATTO488 conjugate prepared from purified actin. |
| Protein concentration | Actin determined by direct UV absorbance at A290 nm; 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
References
Modulation of actin dynamics as potential macrophage subtype-targeting anti-tumour strategy.
Pergola C, Schubert K, Pace S, et al. Scientific Reports. 2017;7:41434. doi:10.1038/srep41434.
Handbook Actin-Toolkit TIRFM
(Web version only for demonstration purpose!
A print version comes with the kit)
Flyer Actin-Toolkits
Material and Safety Data Sheet
Protein Sequence on NCBI
