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Actin-Toolkit SPR (alpha skeletal/cardiac muscle actin)
Novel Kinetic and Affinity Analysis of F-Actin Ligands
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.
Applications
• Characterization of binding kinetics and dissociation constants of ligands binding to F-actin
• Analysis of proteins, lipids or pharmacological compounds altering ligand-actin-interaction
Description
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Protein
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Actin (α-skeletal/cardiac muscle Actin)
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Origin
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Skeletal/Cardiac muscle alpha actin, rabbit/bovine
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Molecular mass
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42kDa
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Toolkit description
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Binding of ligands to filamentous actin is used as a functional proof to state the
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biological activity of a ligand and characterizes the ligand to be an actin binding
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protein. Furthermore, the specific actin binding strongly indicates that the ligand is
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correctly folded and structurally intact.
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The Actin-Toolkit SPR provides a unique method to precisely analyze ligand-F-actin
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interactions by Surface Plasmon Resonance techniques on Streptavidin chips. To
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overcome the general problem in SPR - which is that the actin filaments clog the
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capillaries of the device - short actinfilaments with AnkerproteinTM on the fast
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polymerizing barbed ends are used for for coating. The actin filaments protrude
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from the surface of the chip and permit the introduction of ligands. After the actin
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filaments become attached to the surface of the chip, ligands can be introduced
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to investigate binding to F-actin.
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The Actin-Toolkit SPR was developed to analyze the actin-binding properties of
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ligands (native or recombinant proteins, recombinant fragments etc.) and to support
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the detection even of cryptic actin-binding sites. Further it is suitable to investigate
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effects of pharmacological compounds on ligand-actin complexes, as well as
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further proteins, lipids etc. to ligand-actin complexes.
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The kit contains rabbit skeletal muscle actin (Cat. # 8010-03), AnkerproteinTM, FAB-
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P, GAP-EP (SPR actin buffers for F-and G-actin). A handbook introduces and
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E guides through the experimental work with actin in SPR.
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Please note that the user requires Streptavidin coated chips. The setup for the
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compounds to investigate has to be determined individually.
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Purity
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>99% by scanning densitometry
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Properties
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Form
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Lyophilized, ready-to-use
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Kit contents
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4 x 250 µg Actin (lyophilized powder)
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4 x 10 µg AnkerproteinTM (lyophilized powder)
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1 x PolyMix (actin polymerization buffer, 10x)
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2 x 250 ml FAB-EP, liquid
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1 x 250 ml GAB-EP, liquid
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1 x Toolkit Handbook SPR
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Actin Buffers
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FAB-P: 10mM Imidazol-Cl pH 7.4, 0.4mM ATP, 2mM MgCl2 and 0.0005% v/v
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Surfactant P20.
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GAB-P: 2mM Tris-Cl pH 8.2, 0.4mM ATP, 0.5mM DTT, 0.1mM CaCl2 and
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0.0005% v/v Surfactant P20.
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Purification notes
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Purified from skeletal muscle, rabbit/cardiac muscle, bovine; GPC.
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Protein concentration
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Determined by the Biuret method
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Storage instructions
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We guarantee stable performance of the kit components for 6 months when stored
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at -70°C upon arrival. Solubilized G-actin is kept on ice and should be used
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within 1 week. Avoid refreezing.
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Shipping conditions
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At ambient temperature. Upon delivery store at -70°C.
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Remarks
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For Use in Research only. Not for Use in Human or Veterinary Diagnostical or
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Therapeutical Applications.
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Related products
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For dilution of G-actin or exchange of ligand buffer into actin compatible buffer,
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MonoMix (Cat. #: 5100-0*) may be used. In the absence of nucleators, the
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polymerization of G-actin is initiated by PolyMix (Cat. #: 5000-0*).
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