Glyoxalase I / GLO1 (1-184) Human Protein

Specifications

Product Data
Species	Human
Expression Host	E. coli
Expression cDNA Clone or AA Sequence	Protein Sequence (showhide) MAEPQPPSGG LTDEAALSCC SDADPSTKDF LLQQTMLRVK DPKKSLDFYT RVLGMTLIQK CDFPIMKFSL YFLAYEDKND IPKEKDEKIA WALSRKATLE LTHNWGTEDD ETQSYHNGNS DPRGFGHIGI AVPDVYSACK RFEELGVKFV KKPDDGKMKG LAFIQDPDGY WIEILNPNKM ATLM
Predicted MW	20.7 kDa
Concentration	lot specific
Purity	>90% by SDS - PAGE
Buffer	Presentation State: Purified State: Liquid purified protein Buffer System: 20 mM Tris-HCl buffer (pH 8.0) containing 1 mM DTT, 10% glycerol
Preparation	Liquid purified protein
Protein Description	Recombinant human GLO1 protein was expressed in E.coli and purified by using conventional chromatography techniques.
Storage	Store undiluted at 2-8°C for up to two weeks or (in aliquots) at -20°C or -70°C for longer. Avoid repeated freezing and thawing.
Stability	Shelf life: one year from despatch.
Bioactivity	Biological: >1.7 units/ml (please enquire for specific batch value). One unit will form 1.0 umol of S-lactoylglutathione from methylglyoxal and reduced glutathione per minute at pH 7.5, at 25°C. Specific activity was expressed in units/mg of protein. Activity Assay Final assay concentrations: In 1.5ml reaction mix, the final concentrations are 79 mM potassium phosphate, 0.033%(w/v) reduced glutathine, 0.003% (w/v) bovine serum albumin, 0.033% methylglyoxal (w/v) and recombinant glyoxalase I (0.5ug, 1ug, 2ug). 1. Prepare 1.45ml reaction buffer without recombinant protein into suitable container (for each protein concentration).  2. Equilibrate protein-free reaction buffer to 25C and monitor at A240nm until the value is constant, using a spectrophotometer.  3. Dilute various concentrations of recombinant glyoxalase I (0.5ug, 1ug, 2ug) in 50ul of 10mM potassium phosphate buffer with 0.1% (w/v) BSA, pH 7.4.  4. Add 50ul of recombinant-glyoxalase I to 1.45ml reaction buffer.  5. Immediately mix by inversion and record the increase at A240nm for 5 minutes.
Reference Data
RefSeq	NP_006699
Locus ID	2739
UniProt ID	Q04760, X5DNM4
Cytogenetics	6p21.2
Synonyms	GLOD1; GLYI; HEL-S-74
Summary	The enzyme encoded by this gene is responsible for the catalysis and formation of S-lactoyl-glutathione from methylglyoxal condensation and reduced glutatione. Glyoxalase I is linked to HLA and is localized to 6p21.3-p21.1, between HLA and the centromere. [provided by RefSeq, Jul 2008]
Protein Pathways	Pyruvate metabolism

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