Provides Hot start Enhances PCR specificity and sensitivity Less background Works for any full-lengh Taq

DESCRIPTION
The Taq Start Monoclonal Antibodies were derived from a hybridoma (fusion of mouse myeloma cell and the cells after mouse immunization with Taq DNA Polymerase). Taq Start Monoclonal Antibody is mouse IgG 2b isotype.
When mixed with Taq DNA Polymerase, it provides an antibody-mediated hot start that enhances the specificity, sensitivity, and convenience of PCR. Inhibition is effective during reaction assembly at ambient temperature, and is completely reversed when thermal cycling begins, with no other effect on PCR conditions. Taq Start Monoclonal Antibody is effective with a variety of commercially available Taq DNA polymerases (native or recombinant). The use of Taq Start Monoclonal Antibody significantly improves the specificity of PCR amplification what is especially important for PCR-based diagnostics.

APPLICATION

Hot Start PCR
PCR diagnostics

CONCENTRATION
4 mg/ml (=10 u/µl)

UNIT DEFINITION
One unit is defined as the amount of Taq Start Monoclonal Antibody required to block 50% of activity of 1 µg of Taq DNA Polymerase at 37°C.

STORAGE BUFFER
10 mM Tris-HCl (pH 7.0 at 22°C), 50 mM KCl, 0.1 mM EDTA, 50% glycerol

STORAGE TEMPERATURE
-20°C

REACTION BUFFER
The Taq Start Monoclonal Antibody reaction buffer is the same buffer used for the thermostable DNA polymerase.

PURITY
More than 95% in SDS electrophoresis in 15% PAAG.

ASSOCIATED ACTIVITIES
No conversion to the covalently closed circular DNA to the nicked or linear form was observed after incubation of 1 µg of pUC19 with antibodies in final concentration of 6 u/µl in 20 µl of reaction mixture containing 25 mM Tris-HCl (pH 7.9), 100 mM NaCl, 10 mM MgCl₂ after 16 hours at 37°C.

PROTOCOL
The amount of antibodies required for Taq polymerase activity inhibition depends not on the units of the enzyme, rather the amount of Taq polymerase as protein! The ratio units/mg of Taq polymerase varies strongly from preparation to preparation of the different producers. We recommend to use 2 µl of antibody per 8 µl (40 units) of BioThermTM Taq DNA polymerase.

U/mg RATIO
2500 units of specific activity are equal to 1 mg of antibodies