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Isopeptidase-Fluorogenic
Assays
ZediXcite 330/418 Fluorogenic TG2-Assay Kit
Isopeptidase-activity based TG2 assay (330/418 nm) Art. No.
F014
Synonym
ZediXcite 330/418 Fluorogenic TG2-Assay Kit
Description
Isopeptidase-activity based ready to use kit for fluorescent measurement optimized for tissue transglutaminase (TG2, transglutaminase 2). TG2 cleaves a dark quenching molecule (2,4 dinitrophenyl) from the side chain of a modified peptide incorporating glycine methyl ester. Subsequently, the fluorescence of an N-terminal coupled fluorophore (2-amino benzoic acid, 2-Abz) increases and can be monitored online (excitation wavelength 313-330 nm; emission wavelength 418 nm). The assay is suitable for research purposes, drug discovery programs, and quality assurance.
Assay principle
TISSUE TRANSGLUTAMINASE (TG2) cleaves a dark quenching molecule from the side chain of TG2 specific peptide A163 incorporating glycine methyl ester. Subsequently, the fluorescence of an N-terminal coupled dye increases and can be monitored (excitation wavelength 330 nm; emission wavelength 418 nm).
Reagents in the kit
(1) SUBSTRATE REAGENT (SR): 2 x 23 μL TG2 specific peptide A163 (DMSO solution).
(2) BUFFER REAGENT (BR): 2 x 18 mL TRIS buffer pH 7.5 containing calcium chloride, sodium chloride, glycine methyl ester (lyophilizate).
(3) DILUTION BUFFER (DB): 1 x 18 mL TRIS buffer pH 8.1 containing sodium chloride, EDTA, DTT (lyophilizate).
(2) BUFFER REAGENT (BR): 2 x 18 mL TRIS buffer pH 7.5 containing calcium chloride, sodium chloride, glycine methyl ester (lyophilizate).
(3) DILUTION BUFFER (DB): 1 x 18 mL TRIS buffer pH 8.1 containing sodium chloride, EDTA, DTT (lyophilizate).
Equipment
The ZediXcite Fluorogenic TG2-Assay Kit (F014) can be used in fluorescence microplate readers using microplates as well as in standard fluorescence spectrophotometers with cuvettes. Refer to the instructions of the manufacturer.
Application
Determination of TISSUE TRANSGLUTAMINASE (TG2) activity, specific and sensitive. Suitable for measurements of 1.25 nM to 10 nM (100 ng/mL to 780 ng/mL) of TG2.
Storage
Store at 2-8 °C
Reference(s)
Parameswaran, K.N. et al. J. Biol. Chem. 1997, 272, 10311.
Oertel, K. et al. Anal. Biochem. 2007, 367, 152.
Oertel, K. et al. Anal. Biochem. 2007, 367, 152.
Note
INTENDED FOR RESEARCH USE ONLY, NOT FOR USE IN HUMAN, THERAPEUTIC OR DIAGNOSTIC APPLICATIONS.
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