tebu-bio is a European company specialised in providing innovative reagents and laboratory services in Life Sciences
X

Need an answer now?

Give us a call

Leave a message, we'll get back to you





Actin Polymerization Kit

See the related products

Introduction

The Actin Polymerization Biochem Kit™ is based on the enhanced fluorescence of pyrene conjugated actin that occurs during polymerization. The enhanced fluorescence that occurs when pyrene G-actin (monomer) forms pyrene F-actin can be measured in a fluorimeter to follow polymerization over time. Also, by using preformed pyrene F-actin, it is possible to follow depolymerization. Both cell/tissue extracts and purified proteins can be added to the reaction mixture to identify their effect on actin polymerization. The components of the kit can also be used separately for other actin based assays such as a spin-down assays to detect F-actin binding proteins (see also Cat nr. BK001) or size exclusion chromatography to identify G-actin binding proteins.

 

While this kit comes with pyrene labeled skeletal muscle actin, it can also be used to study polymerization of other types of actin such as non-muscle actin (Cat nr. APHL99) or cardiac actin (Cat nr. AD99). Polymerization assays with these actins can be performed using a 10:1 ratio between the actin you want to study and the included pyrene actin

Kit contents

The kit contains enough materials for 30-100 assays depending on assay volume. The following reagents are included:

  1. 5 x 1 mg Pyrene labeled actin (Cat nr AP05). 
  2. General Actin Buffer (Cat nr BSA01).
  3. Actin Polymerization Buffer (Cat nr BSA02).
  4. ATP 100mM (Cat nr BSA04).
  5. Tris-HCl pH 7.5, 100 mM
  6. Manual with detailed protocols and extensive troubleshooting guide.

Equipment needed

  1. Fluorescence spectrophotometer (cuvette or 96-well plate) with 4-10 nm bandwidth at 365 nm excitation wavelength, and 4-10 nm bandwidth at 407 nm emission wavelength.
  2. Small capacity (100-1000 µl) fluorescence spectrophotometer cuvette or 96-well plate.

Example results

The Actin Polymerization Biochem Kit™ was used to study the effects of Arp2/3 (Cat nr RP01) and the VCA domain of WASP on actin polymerization rates. The Arp2/3 complex is an actin filament nucleator but has low nucleating/polymerizing activity on its own. The VCA domain of WASP is an activator of the Arp2/3 complex. Hence, when the Arp2/3 complex is mixed with the WASP VCA domain, these two exert a potent actin polymerizing activity (Fig. 1 in the Gallery, Related Downloads section).

Product Uses Include


See the related products