This Human ECE2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of ECE2 protein (Cat: 10207-H07H) from the overexpression lysate was verified.
A DNA sequence encoding the ectodomain of human endothelin converting enzyme 2 isoform A (NP_055508.3) (Gly 199-Trp 883) was fused with a polyhistidine tag at the N-terminus.
The recombinant human ECE2 consists of 700 amino acids with the predicted molecular mass of 80.2 kDa. As a result of glycosylation, rhECE2 migrates as an approximately 120-125 kDa band in SDS-PAGE under reducing conditions.
Human ECE2 HEK293 Overexpression Lysate: 用法
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human ECE2 HEK293 Overexpression Lysate: 別名
Human KIAA0604 Overexpression Lysate; Human UNQ403/PRO740 Overexpression Lysate
Endothelin-converting enzyme 2, also known as ECE-2, is a metalloprotease that possesses many properties consistent with it being a neuropeptide-processing enzyme. Endothelin-converting enzymes (ECEs) are the key enzymes in the endothelin (ET) biosynthesis that catalyze the conversion of big ET, the biologically inactive precursor of mature ET. Two enzymes, termed ECE-1 and ECE-2, have been molecularly identified. ECE-2 is found primarily in neural tissues, with high levels of expression in midbrain, cerebellum, hypothalamus, frontal cortex and spinal cord and moderate levels in hippocampus and striatum. ECE-2 is strongly down-regulated in inferior parietal lobe from Alzheimer disease patients (at protein level). ECE-2 converts big endothelin-1 to endothelin-1. It is involved in the processing of various neuroendocrine peptides, including neurotensin, angiotensin I, substance P, proenkephalin-derived peptides, and prodynorphin-derived peptides. ECE-2 may limit beta-amyloid peptide accumulation in brain. It may also have methyltransferase activity. A comparison of residues around the cleavage site revealed that ECE-2 exhibits a unique cleavage site selectivity that is related to but distinct from that of ECE-1.
endothelin converting enzyme 2
Lorenzo M.-N., et al.,(2001), Human endothelin converting enzyme-2 (ECE2): characterization of mRNA species and chromosomal localization. Biochim. Biophys. Acta 1522:46-52.
Muzny D.M., et al., (2006), The DNA sequence, annotation and analysis of human chromosome 3.Nature 440:1194-1198.
Nagase T., et al.,(1998), Prediction of the coding sequences of unidentified human genes. IX. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro.DNA Res. 5:31-39.