Actin-Toolkit TIRFM (ATTO647-Actin)

Protein

ATTO647-Actin (α-Actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

The Actin-Toolkit TIRFM was developed to analyze actin dynamics by TIRF microscopy or other fluorescence based single molecule imaging techniques. The Toolkit contains all components required for TIRFM analyses and enables the user to start TIRFM experiments straight ahead. 

It includes ATTO647-Actin (λex 647nm, λem 689nm) with a DOL of 0.3 as demanded for TIRFM analysis. The Toolkit contains NEM-Myosin II as a tethering protein, and all components required for flow chamber TIRF assays, including FluMaXx - an efficient oxygen scavenger system - to protect actin filament from fragmentation during analyses. A detailed description of the flow chamber assembly and TIRFM actin polymerization assays is provided with the handbook. 

In combination with the actin buffers contained in the kit, error-free handling is guaranteed. The kit contains 5x100 µg of α-skeletal muscle ATTO647-Actin conjugates and buffers to keep the actin functional.

Purity

>99% by scanning densitometry.

Form

Lyophilized and in solution (see kit content).

Kit content

5x100 µg ATTO647-Actin (lyophilized powder);
1x100 µg NEM-Myosin II (lyophilized powder);
3x1 ml Passivation solution (5% BSA, fatty acid free);
1x1 ml PolyMix (lyophilized F-actin ATP-buffer);
1x50 ml MonoMix (lyophilized G-actin ATP-buffer);
1x10 ml HSB 5x stock (solution,Myosin Buffer, pH 7.0);
1x0.5 ml ME-Buffer (100x Mg-Exchange buffer);
1x Toolkit Handbook TIRFM;
 

+ FluMaXx
Oxygen Scavenger System for single molecule imaging 

(Cat. #: 5161-01)

Content

5x250 µl FluMaXx (2x stock, lyophilized)   
Contains: Glucose oxidase, catalase, imidazole pH 7.4, KCl, EGTA, MgCl2, glucose, cryoprotectants (proprietary formula);
1x500 µl ATP/DTT Mix (10x stock, lyophilized)
Contains: ATP, DTT 
2x1.5 ml Methylcellulose stock solution (0.5% in Milli-Q™ water)

 

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.

Related products

Actin Binding Proteins

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)