Actin-Toolkit Fluorescence Microscopy (ATTO532-Actin, skeletal muscle actin)

Protein

ATTO532-Actin (α-Actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

The Actin-Toolkit Fluorescence Microscopy was developed to analyze and demonstrate the effect of ligands on actin polymerization, or ligand-induced formation of actin bundles and networks. In the assays,  ATTO532-Actin (λex 532nm, λem 553nm) is mixed with the ligand to observe the effects on either G- or F-actin. In contrast to staining with phalloidin conjugates, which can interfere with ligand binding sites on the actin molecule, no such observations were made with ATTO labeled actin. ATTO-Actins are validated to possess the polymerization properties of native actin.

The Toolkit further contains unlabeled actin (0.5mg α-skeletal muscle actin). A mixture of both excludes possible interference of the fluorescent actin conjugates on ligand binding sites. Vinculin tail - a V8 protease cleavage product of vinculin -  is an efficient actin bundling vinculin domain, is supplied with the Toolkitas a bundling reference.

Several standardized experiments are described step-by-step in the handbook supplied with the kit. In combination with the actin bufferscontained in the kit, error-free handling is guaranteed. The kit contains 4x100µg of α-skeletal muscle ATTO532-Actin conjugates and buffers to keep the actin functional.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

4x100µg ATTO532-Actin (lyophilized powder);
1x0.5mg α-skeletal muscle actin (lyophilized powder);
1x50µg Vinculin tail;
5x50ml MonoMix (lyophilized G-actin ATP-buffer);
1x1.0ml PolyMix 10x stock (lyophilized F-actin ATP-buffer, pH 7.4);
1x0.5ml 1M MgCl2;
1x Toolkit Handbook Fluorescence Microscopy;
10x Light protective tubes & tags;

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)