Photometric Activity Assay

Blood Coagulation Factor XIII-Microassay Kit, colorimetric

1 Kit 415 € available

Art. No. T114
Synonym FXIII-CovTest
Assay principle The Blood Coagulation Factor XIII-Microassay Kit uses biotinylated F11 peptide (BiotinpepF11) as the first substrate (amine-acceptor/acyl-donor) and spermine as second substrate (amine-donor/acyl-acceptor).
Usually samples contain the pro-enzyme (FXIII) and a pre-treatment with thrombin (T056) to obtain activated FXIII (FXIIIa) could be necessary to measure the activity using Blood Coagulation Factor XIII-Microassay Kit. After this activation step, samples suspected of containing activated FXIIIa are incubated with calcium, dithiothreitol (DTT), and Biotin-pepF11 in the wells of microtiter plates to which spermine had been covalently coupled, previously. In the presence of FXIII spermine is incorporated into the gamma-carboxamide of the glutaminyl residue of the biotin-pepF11 to form a biotin-pepF11-gamma-glutamyl spermine.
The system is coupled to Streptavidin labelled peroxidase (SAv-HRP). SAv-HRP is revealed using H2O2 as HRP substrate and tetramethyl benzidine as electron acceptor (chromogen).
Reagents in the kit Microtiter strips with covalently bound spermine
EDTA (Negative Control)
Reaction Buffer (Biotin-pepF11/CaCl2)
Enzyme Tracer (SAv-HRP)
Wash Buffer 10X
Diluent Buffer 10X
HRP Substrate
Blocking Reagent
Equipment Microplate reader capable of measuring absorbance at 450 nm.
Pipettes, multi-channel pipette, and pipette tips.
Ultra-pure or deionized water.
Squirt bottle, manifold dispenser, or automated microplate washer.
Application Colorimetric measurement of transglutaminase activity, optimized for blood coagulation factor XIII
Storage The kit is shipped on ice. Upon arrival, the Reaction Buffer and Enzyme Tracer should be stored at -20°C. All the other components of the kit should be kept at +4°C.
When stored properly, these stock solutions are stable for at least 24 months.
Note Kit produced and Developed by CovalAb, France



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