Lactoylglutathione lyase, also known as GLO1, belongs to the glyoxalaseIfamily. Glyoxalase I is responsible for the catalysis and formation of S-lactoyl-glutathione from methylglyoxal condensation and reduced glutathione.This enzyme is ubiquitously expressed and is also present in many tumor cell lines, in which its concentration is often upregulated. Source: Recombinant protein corresponding to aa1-184 from human GLO1, expressed in E. coli. Molecular Weight: ~20.7kD (184aa) Specific Activity: >400units/mg, one unit will form 1umol of S-lactoylgutathione from methylglyoxal and reduced glutathione/minute at pH 6.5 at 25C. Activity Assay: 1. Prepare 180ul assay buffer into a suitable container: The concentrations are 88mM potassium phosphate, pH 7.5, 1.1mM reduced glutathione, 3.3mM methylglyoxal. 2. Equilibrate to 25C and monitor the A240nm until value is constant using a spectrophotometer. 3. Add 20ul of recombinant GLO1 protein 0.025ug in assay buffer. 4. Record the increase in A240nm for 5 minutes. Amino Acid Sequence: ARPCIPKSFG YSSVVCVCNA TYCDSFDPPT FPALGTFSRY ESTRSGRRME LSMGPIQANH TGTGLLLTLQ PEQKFQKVKG FGGAMTDAAA LNILALSPPA QNLLLKSYFS EEGIGYNIIR VPMASCDFSI RTYTYADTPD DFQLHNFSLP EEDTKLKIPL IHRALQLAQR PVSLLASPWT SPTWLKTNGA VNGKGSLKGQ PGDIYHQTWA RYFVKFLDAY AEHKLQFWAV TAENEPSAGL LSGYPFQCLG FTPEHQRDFI ARDLGPTLAN STHHNVRLLM LDDQRLLLPH WAKVVLTDPE AAKYVHGIAV HWYLDFLAPA KATLGETHRL FPNTMLFASE ACVGSKFWEQ SVRLGSWDRG MQYSHSIITN LLYHVVGWTD WNLALNPEGG PNWVRNFVDS PIIVDITKDT FYKQPMFYHL GHFSKFIPEG SQRVGLVASQ KNDLDAVALM HPDGSAVVVV LNRSSKDVPL TIKDPAVGFL ETISPGYSIH TYLWRRQHHH HHH Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
