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작성일 : 19-05-30
[Hypermol EK] Novel Kinetic & Affinity Analysis of F-Actin Ligands
Actin-Toolkit SPR (alpha skeletal/cardiac muscle actin)

 

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.

 


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

 Protein

 

Actin (α-skeletal/cardiac muscle Actin)

 Origin

 

Skeletal/Cardiac muscle alpha actin, rabbit/bovine

 Molecular mass

 

42kDa

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.

 Purity

 

>99% by scanning densitometry

 

Properties

 Form

 

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.

 Remarks

 

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

Size

8090-01

Actin-Toolkit SPR (alpha skeletal muscle actin)

kit

8900-01

Actin-Toolkit SPR (alpha cardiac muscle actin)

kit

 

 

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