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Actin-Tookit Fluorometry (alpha skeletal muscle alpha actin, pyrene-actin)

Cat. #:
8030-01
Shipping:
FedEx Express (Global) FedEx Express (Global) (shipping info)
Units:
1x kit


Actin-Toolkit Fluorometry
& Pyrene Assays

Rabbit Skeletal Muscle α-Actin


Cat. # 8030-01 

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Applications

  • Kinetic analysis of purified ligands, cell or tissue extracts on polymerization or depolymerization of actin;
  • Determination of sensitivity to divalent cations or pharmacological compounds during polymerization or depolymerization; 
  • Measurement of F-actin severing effects of purified ligands, cell or tissue extracts at steady-state;
 
 

Description

   

Protein

Pyrene Actin (α-Actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

The Actin-Toolkit Fluorometry is used to determine the polymerization and depolymerization reaction of actin by measuring the fluorescence of pyrenyl conjugatedactin. By this means it also allows to analyze the interaction of actin with ligands or other components effecting the polymerization or depolymerization. The fluorescence of Pyrene Actin is maximally increased by a factor of 22 upon polymerization. The Actin-Toolkit Fluorometry is suitable to measure the effect on actin polymerization of purified ligands, cell or tissue extracts.  

The Actin-Toolkit Fluorometry all-in-one kit contains components to measure the effects of native or recombinant proteins or fragments, lipids or other ligands on actin dynamics. The kit contains 8x1mg pure rabbit skeletal muscle actin mixed with 10% pyrenyl actin (dye/protein molar ratio = 1:10) labelled at Cys 374 with N-(1-pyrene) iodoacetamide buffers to keep actin functional and to vary e.g. the Ca++-concentration. 

A handbook with a detailed description of experimental setups guides the user for safe, error-free handling of pyrenyl actin. We guarantee stable performance of the kit components for 6 months when stored at -70°C upon arrival. Experienced users may supplement this kit with 100% Pyrene Actin (dye/protein molar ratio = 1:1), to increase the signal by using for instance 20% pyrene actin in the assays. 

Purity

>99% by scanning densitometry.



Properties

   

Form

Lyophilized, ready-to-use.

Kit content

8x1.0mg Pyrene Actin 10% (lyophilized powder);
1x40µg Gelsolin (mammalian);
1x1000ml ATP-Actin Buffer, supplied as components; 
1x1.0ml PolyMix (10x)
1x0.5ml CaCl2 (100mM);
1x Toolkit Handbook Fluorometry;
8x Black 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)




Further Information


Material and Safety Data Sheet


Protein Sequence on NCBI






 

 
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