Novel Kinetic & Affinity Analysis of F-Actin Ligands



Actin-Toolkit SPR (alpha skeletal/cardiac muscle actin)

Novel Kinetic and Affinity Analysis of F-Actin Ligands

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The Actin-Toolkit SPR is a unique method to analyze ligand interactions with actin filaments. The kit is

available with α-actin (rabbit skeletal muscle) and bovine α-cardiac actin.  

The analysis of proteins, lipids or pharmacological compounds to analyze kinetics and affinity of ligands

specifically binding to F-Actin.



• Characterization of binding kinetics and dissociation constants of ligands binding to F-actin

• Analysis of proteins, lipids or pharmacological compounds altering ligand-actin-interaction





Actin (α-skeletal/cardiac muscle Actin)



Skeletal/Cardiac muscle alpha actin, rabbit/bovine

 Molecular mass



Toolkit description


Binding of ligands to filamentous actin is used as a functional proof to state the



biological activity of a ligand and characterizes the ligand to be an actin binding



protein. Furthermore, the specific actin binding strongly indicates that the ligand is



correctly folded and structurally intact.



The Actin-Toolkit SPR provides a unique method to precisely analyze ligand-F-actin



interactions by Surface Plasmon Resonance techniques on Streptavidin chips. To



overcome the general problem in SPR - which is that the actin filaments clog the



capillaries of the device - short actinfilaments with AnkerproteinTM on the fast



polymerizing barbed ends are used for for coating. The actin filaments protrude



from the surface of the chip and permit the introduction of ligands. After the actin



filaments become attached to the surface of the chip, ligands can be introduced



to investigate binding to F-actin.



The Actin-Toolkit SPR was developed to analyze the actin-binding properties of



ligands (native or recombinant proteins, recombinant fragments etc.) and to support



the detection even of cryptic actin-binding sites. Further it is suitable to investigate



effects of pharmacological compounds on ligand-actin complexes, as well as



further proteins, lipids etc. to ligand-actin complexes.



The kit contains rabbit skeletal muscle actin (Cat. # 8010-03), AnkerproteinTM, FAB-



P, GAP-EP (SPR actin buffers for F-and G-actin). A handbook introduces and



E guides through the experimental work with actin in SPR.



Please note that the user requires Streptavidin coated chips. The setup for the



compounds to investigate has to be determined individually.



>99% by scanning densitometry





Lyophilized, ready-to-use

 Kit contents


4 x 250 µg Actin (lyophilized powder)



4 x 10 µg AnkerproteinTM (lyophilized powder)



1 x PolyMix (actin polymerization buffer, 10x)



2 x 250 ml FAB-EP, liquid



1 x 250 ml GAB-EP, liquid



1 x Toolkit Handbook SPR

 Actin Buffers


FAB-P: 10mM Imidazol-Cl pH 7.4, 0.4mM ATP, 2mM MgCl2 and 0.0005% v/v



Surfactant P20.



GAB-P: 2mM Tris-Cl pH 8.2, 0.4mM ATP, 0.5mM DTT, 0.1mM CaCl2 and



0.0005% v/v Surfactant P20.

 Purification notes


Purified from skeletal muscle, rabbit/cardiac muscle, bovine; 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.



For Use in Research only. Not for Use in Human or Veterinary Diagnostical or



Therapeutical Applications.

 Related products


For dilution of G-actin or exchange of ligand buffer into actin compatible buffer,



MonoMix (Cat. #: 5100-0*) may be used. In the absence of nucleators, the



polymerization of G-actin is initiated by PolyMix (Cat. #: 5000-0*).



Ordering informations

Catalog No.

Product Name



Actin-Toolkit SPR (alpha skeletal muscle actin)



Actin-Toolkit SPR (alpha cardiac muscle actin)




▣ 관련 페이지 ; HYPERMOL EK





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