Celiac disease antigens

Human epidermal transglutaminase

(TG3, recombinantly produced in insect cells)
Quantity Unit Price Status
200 µg 400 € Available
Documents
Art. No.
T024
Synonym
TG3, TGase 3, epidermal protein-glutamine-γ-glutamyltransferase, TGE
Background info
This product replaces product replaces product number T124.
Molecular Weight
78 kDa
Source
Recombinantly produced in insect cells
Activation
His6-rhTG3 is a Ca2+-dependent enzyme.
10 μl His6-rhTG3 (1 μg/μl)
35 μl 50 mM Tris-HCl pH 8
5 μl Dispase I (Roche) (0.2 μg/μl)
Incubate for 20 min at 30°C
Add 10 mM Ca2+ when measuring His6-rhTG3 activity.
Specific Activity
> 1000 U/mg [Activity is determined by measuring the rate of fluorescence enhancement after His6-rhTG3-catalyzed monodansylcadaverine-incorporation into N,N-dimethylated casein according to Lorand et al., Anal. Biochem. 44 (221-231)].
1 U is defined as the increase in fluorescence intensity of 1 a.u./min (measured on a Cary eclipse fluorescence spectrophotometer, Varian; λex = 332 nm, λem = 500 nm; band filter = 5 nm; detector strength = 600 V; temperature = 37°C, assay volume = 1 ml).
Appearance
White lyophilized solid.
Description
His6-rhTG3 is based on the TGM3-allele from clone DKFZp686J0716 (isolated from different tissues), corrected by the insertion of the missing T at Position 435. It is N-terminally fused to a hexahistidine-tag resulting in the encoded N-terminal amino acid sequence MHHHHHHAALGV… . His6-rhTG3 is produced in insect cells and purified by ion metal chelating and ion exchange chromatography to more than 95 % purity.
Reagents
The Transglutaminase is lyophilized from 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA and 1 mM DTT. Sample contains maltodextrin.
Reconstitution
Add the volume of water specified in the certificate of analysis under aliquotation to the vial of lyophilized powder. Rotate vial gently until solid dissolves. After reconstitution the solution should be stored frozen in working aliquots. For short term storage keep cooled on ice.
Application
His6-rhTG3 catalyzes acyl transfer reactions from glutamine residues in proteins or peptides to primary amines, e. g. the formation of ε-(γ-glutamyl) lysine bonds between proteins by transferring the acyl group of a peptide-bound glutamine residue to the primary amino group of a peptide-bound lysine residue. His6-rhTG3 may also be used for immunoprecipitation and the detection of TG3 autoantibodies.
Storage
Storage for several months is possible at ≤ - 20°C.
Delivery is possible at ambient temperature
Reference(s)
Hietikko et al., Acta. Derm. Venereol. 2018, 98:366-72
Yamane et al., FEBS J. 2010, 277:3564-74
Note
INTENDED FOR RESEARCH USE ONLY, NOT FOR USE IN HUMAN, THERAPEUTIC OR DIAGNOSTIC APPLICATIONS.

This product replaces TG3, recombinantly produced in E. coli (T012).

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