PDIA4, also known as ERP72, is an endoplasmic reticulum luminal protein that is both a stress protein and a member of the protein disulfide isomerase family of proteins. It is involved in the catalysis of protein- S-S-bond rearrangement. PDIA3 and PDIA4 act as proteases, protein disulfide isomerases, phospholipases or a combination of these. Source: Recombinant protein corresponding to aa21-645 from human PDIA4, fused to His-tag at N-terminal expressed in E. coli. Molecular Weight: ~72.9kD (646aa) Specific Activity: >10 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 PDIA4 to 800ug/ml in 0.1% BSA. Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MVAGAEGPDE DSSNRENAIE DEEEEEEEDD DEEEDDLEVK EENGVLVLND ANFDNFVADK DTVLLEFYAP WCGHCKQFAP EYEKIANILK DKDPPIPVAK IDATSASVLA SRFDVSGYPT IKILKKGQAV DYEGSRTQEE IVAKVREVSQ PDWTPPPEVT LVLTKENFDE VVNDADIILV EFYAPWCGHC KKLAPEYEKA AKELSKRSPP IPLAKVDATA ETDLAKRFDV SGYPTLKIFR KGRPYDYNGP REKYGIVDYM IEQSGPPSKE ILTLKQVQEF LKDGDDVIII GVFKGESDPA YQQYQDAANN LREDYKFHHT FSTEIAKFLK VSQGQLVVMQ PEKFQSKYEP RSHMMDVQGS TQDSAIKDFV LKYALPLVGH RKVSNDAKRY TRRPLVVVYY SVDFSFDYRA ATQFWRSKVL EVAKDFPEYT FAIADEEDYA GEVKDLGLSE SGEDVNAAIL DESGKKFAME PEEFDSDTLR EFVTAFKKGK LKPVIKSQPV PKNNKGPVKV VVGKTFDSIV MDPKKDVLIE FYAPWCGHCK QLEPVYNSLA KKYKGQKGLV IAKMDATAND VPSDRYKVEG FPTIYFAPSG DKKNPVKFEG GDRDLEHLSK FIEEHATKLS RTKEEL 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.
