Angiotensin Converting Enzyme 2, Recombinant, Human, Control Peptide (ACE-2, ACEH)

Molecular Biology / MB-Enzymes, ACE

-20°C

Supplied as a lyophilized powder from PBS, 0.05% sodium azide.

Western Blot Positive Control for A2295-01R (affinity purified antibody). Human ACE-2 (1-740aa, glycosylated secreted protein of MW ~120kD, predicted MW85kD) with His tag was expressed in mouse myeloma cell line NS0 and purified >90% (SDS-PGE).

Renin-Angiotensin System (RAS) is a crtical regulator of blood pressure homeostasis. The protease renin cleaves angiotensinogen into inactive decameric peptide angiotensin-I (Ang-I). Angiotensin-converting enzyme (ACE) then cleaves C-terminal dipeptide from Ang-I to form an active octamer angiotensin-II (Ang-II), which can contribute to hypertension by promoting vascular smooth muscle vasocontriction and renal tubule sodium reabsorption. ACE can also cleave many other small peptides including the vasodialating peptide bradykinin into inactive fragment, cleave Alzheimer amyloid beta-peptide (Abeta), retard Abeta aggregation, deposition and fibril formation. ACE mutant mice display spontaneous hypotension, partial male infertility and kidney malformations. ACE is found in somatic (s-ACE) and testicular/germinal (t-ACE) isoforms. The products of renin and ACE catalysis, namely Ang1-10 and Ang1-8 can also be by another peptidase, ACE-2 to Ang1-9 and Ang1-7, respectively. ACE-2 and ACE (s-ACE and t-ACE) are made as transmembrane (TM) proteins. These enzymes also exist as soluble, truncated forms lacking the TM and cytosolic domains.

ACE-2 (also known as ACE-2 and ACE homolog, ACEH) gene has been mapped at human chromosome Xp22. ACE-2 enzymes from human (805aa) and mouse (798aa) are single chain proteins with 40% seq homology to N- and C-terminal domains of ACE. However, in contrast to s-ACE which consists of two catalytic sites, ACE-2 contains only one active site. Unlike s-ACE and t-ACE which are dipeptidyl-carboxypeptidases, ACE-2 acts as a carboxypeptidase, cleaving single residue from Ang-I, generating Ang1-9 and a single residue from Ang-II to generate Ang1-7. ACE-2 can cleave angiotensin-I but not bradykinin and the enzyme activity is not inhibited by the ACE inhibitors. This enzyme is expressed highly in heart, kidney and testis and moderately in colon, small intestine and ovary. ACE-2 is an essential regulator of heart fuction because targeted disruption of this enzyme in mice results in severe cardiac contractility defect, increased angiotensin-II levels and upregulation of hypoxia-induced genes in the heart.

Applications:
Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
This preparation is biologically inactive. It is not suitable for ELISA or other applications where native protein is required. It is supplied in 100ul/vial. For Western Blot, heat once and load 10ul/lane and visualize with appropriate antibodies. This preparation is intended for qualitative purpose and not to serve as standard of known concentration. Angiogenin protein is ~14kD. Store frozen in suitable aliquots. Do not freeze, thaw, or heat repeatedly.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile buffer or ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. 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.