Actin-Toolkit ELISA (alpha skeletal muscle actin)

Proteins

Biotin-Actin (biotinylated α-skeletal muscle actin)

Origin

Skeletal muscle alpha actin, rabbit

Molecular mass

42kDa

Toolkit description

The Actin-Toolkit ELISA is used to analyse ligand interactions with G-Actin or F-actin by enzyme linked immunosorbant assay. Purified proteins or other ligands, semi purified ligands or crude extracts can be used for the assays. Thus the Toolkit is suited for quantitative analyses or to determine actin-binding at various concentrations e.g. with crude extracts. 

Two types of ELISA assay are utilized in the assays:

1. Direct ELISA.: the ligand is coated to the surface of the plate and either G-Biotin-actin or F-biotin actin is used to directly interact with the ligand. To detect Biotin-Actin if successfully bound to the coated ligand, either e.g. anti-biotin antibodies (HRP or alk. Phosphatase conjugated) or e.g. Streptavidin  (HRP or alk. Phosphatase conjugated) can be used. 

2. Sandwich ELISA.: the ligand is coated to the surface of the plate and either G-Biotin-actin or F-actin is used to directly interact with the ligand. To detect Actin successfully bound to the coated ligand the anti-actin antibody (clone 2G2) is used to be detected by secondary antibodies (mouse anti IgG or mouse anti IgM - HRP or alk. Phosphatase conjugated) is used. Anti-Actin is an IgM and thus able to efficiently bind to five actin. This results in an amplification of the signal obtained by secondary antibodies.   

The kit contains Biotin-Actin, Actin and monoclonal anti-Actin, buffers for ELISA keeping actin functional and preventing denaturing of actin. A handbook introduces and guides through the experimental work with actin.

Purity

>99% by scanning densitometry.

Form

Lyophilized, ready-to-use.

Kit content

2x100µg Biotin-Actin (lyophilized powder);
4x50ml  G- ATP-buffer (lyophilized);
1x200ml  F- ATP-buffer;
1x100mM ATP (lyophilized);
1x500mM DTT (lyophilized)
1x25µg monoclonal anti-Actin;
1x50mg BSA (fatty-acid free); 
1xToolkit Handbook Actin-Toolkit ELISA;

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

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)