Human ECE2 HEK293 Overexpression Lysate

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Human ECE2 HEK293 Overexpression Lysate: Product Information

Product Description
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-H01H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the ectodomain of human endothelin converting enzyme 2 isoform A (NP_055508.3) (Gly 199-Trp 883) was fused with the Fc region of human IgG1 at the N-terminus.
Molecule Mass
The recombinant human Fc/ECE2 is a disulfide-linked homodimeric protein. The reduced monomer consists of 921 amino acids and predicts a molecular mass of 105 kDa. As a result of glycosylation, the apparent molecular mass of rh ECE2/Fc monomer is approximately 120-130 kDa in SDS-PAGE under reducing conditions.

Human ECE2 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human ECE2 HEK293 Overexpression Lysate: Alternative Names

Human KIAA0604 Overexpression Lysate; Human UNQ403/PRO740 Overexpression Lysate

ECE2 Background Information

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.
Full Name
endothelin converting enzyme 2
References
  • 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.
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