opr0029

Recombinant human epidermal Transglutaminase (His6-rhTG3)

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 hexahistidine-tag resulting in the encoded N-terminal amino acid sequence MHHHHHHAALGV...
His6-rhTG3 is produced in E.coli and purified by ion metal chelating chromatography to more than 95% purity.

Recombinant protein produced in E.coli

His6-rhTG3 catalyzes acyl transfer reactions from glutamine residues in proteins or peptides to primary amines, e. g. the formation of epsilon-(gamma-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.

Purity: >95%
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; lex = 332 nm, lem = 500 nm; band filter = 5 nm; detector strength = 600 V; temperature = 37°C, assay volume = 1 ml)].

White lyophilized solid

The TG is lyophilized from 50 mM Tris-HCl pH 8.0, 5 mM Glutathion

Store working aliquots at a minimum of -20°C
Avoid repeated freezing and thawing
Delivery at ambient temperature is possible

Add the desired volume of H20 to the vial of lyophilized powder. Rotate vial gently until solid dissolves. After reconstitution, the solution should be cooled on ice for short term storage or stored frozen in working aliquots for long term storage.
Activation:
- 10 µl His6-rhTG3 (1 µg/ml)
- 35 µl 50 mM Tris-HCl pH 8
- 5 µl Dispase II (0,2 µg/µl)
Incubate for 20 min at 30°C
Add 10 mM Ca2+ when measuring His6-rhTG3 activity.