PDIA6 is a member of the protein disulfide isomerase (PDI). PDI is an enzyme in the endoplasmic reticulum in eukaryotes or periplasmic space of prokaryotes that catalyzes the formation and breakage of disulfide bonds between cysteine residues within proteins as they fold. PDIA6 function as a chaperone that inhibits aggregation of misfolded proteins. It plays a role in platelet aggregation and activation by agonists such as convulxin, collagen and thrombin Source: Recombinant protein corresponding to aa20-440 from human PDIA6, fused to His-tag at N-terminal expressed in E. coli. Molecular Weight: ~48.5kD (442aa) Specifc Activity: >20 A650/cm/min/mg. Enzymatic activity was confirmed by measuring the aggregation of insulin in the presence of DTT. Activity Assay 1. Prepare a 180ul assay buffer into a suitable container: The concentrations are 0.67mM DTT, insulin 5.3mg, 1.2mM EDTA 2. Dilute insulin 8mg/ml in 3mM EDTA, pH 7.0, and incubate diluted insulin for 5 minutes at 37C. 3. Dilute PDIA6 to 300ug/ml in 0.1%BSA. 4. Mix 40ul diluted PDIA6 and 10mM DTT 10ul. Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MLYSSSDDVI ELTPSNFNRE VIQSDSLWLV EFYAPWCGHC QRLTPEWKKA ATALKDVVKV GAVDADKHHS LGGQYGVQGF PTIKIFGSNK NRPEDYQGGR TGEAIVDAAL SALRQLVKDR LGGRSGGYSS GKQGRSDSSS KKDVIELTDD SFDKNVLDSE DVWMVEFYAP WCGHCKNLEP EWAAAASEVK EQTKGKVKLA AVDATVNQVL ASRYGIRGFP TIKIFQKGES PVDYDGGRTR SDIVSRALDL FSDNAPPPEL LEIINEDIAK RTCEEHQLCV VAVLPHILDT GAAGRNSYLE VLLKLADKYK KKMWGWLWTE AGAQSELETA LGIGGFGYPA MAAINARKMK FALLKGSFSE QGINEFLREL SFGRGSTAPV GGGAFPTIVE REPWDGRDGE LPVEDDIDLS DVELDDLGKD EL 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.
