Nori Human Angiotensin 1-7 ELISA Kit Summary

Alternative names for  angiotensin 1-7: Ang 1-7, Ang (1-7), angiotensin (1-7)

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Angiotensin (1-7) (C₄₁H₆₂N₁₂O₁₁; Molecular weight = 899.02 g/mol; H-Asp–Arg–Val–Tyr–Ile–His–Pro-OH) is an active heptapeptide of the renin–angiotensin system (RAS). In 1988, Santos et al showed that Angiotensin-(1-7) was a main product of the incubation of Angiotensin I with brain micropunches.^[1] In the same year Schiavone et al, reported the first biological effect of this heptapeptide. ^[2] Angiotensin (1-7) is a vasodilator agent that plays important roles in cardiovascular organs, such as heart, blood vessels, and kidneys having functions frequently opposed to those attributed to the major effector component of the RAS, angiotensin II (Ang II).^[3] The polypeptide Ang I can be converted into Ang (1-7) by the actions of neprilysin (NEP)^[4] and thimet oligopeptidase (TOP)^[5] enzymes. Also, Ang II can be hydrolyzed into Ang (1-7) through the actions of angiotensin-converting enzyme 2 (ACE2). Ang (1-7) binds and activates the G-protein coupled receptor Mas receptor^[6] leading to opposite effects of those of Ang II.

Ang (1-7) has been shown to have anti-oxidant and anti-inflammatory effects.^[7][8] Ang (1-7) plays protective roles in cardiomyocytes of spontaneously hypertensive rat by increasing the expression of endothelial and neuronal nitric oxide synthase enzymes leading to augmented production of nitric oxide.^[9] Ultimately, Ang (1-7) evokes anti-arrhythmogenic effects in animal models. In blood vessels, Ang (1-7) induces the release of vasodilators such as prostanoids and nitric oxide. In addition, Ang (1-7) contributes to the beneficial effects of ACE inhibitors and angiotensin II receptor type 1 antagonists.^[10]

References

1. Santos RA, et al. (1988). Hypertension. 11 (2): 125–37.
2. Schiavone MT, et al. (1988). PNAS. 85 (11): 4095–8.
3. ^Santos RA, et al. (2018). Physiol Rev. 1 (98): 505–553.
4. Chappell MC (2016). Am J Physiol. Heart Circulatory Physiol. 310 (2): H137-52.
5. Wilson BA, et al. (2016). Am J Physiol. Renal Physiology. 310 (7): F637–F645.
6. Santos RA, et al. (2003).  PNAS 100 (14): 8258–63.
7. Benter IF, et al. (2008). American Journal of Nephrology. 28 (1): 25–33.
8. El-Hashim AZ, et al. (2012). British Journal of Pharmacology. 166 (6): 1964–76.
9. Zisman LS, et al. (2003). Circulation. 108 (14): 1679–81.
10. Benter IF, et al. (2011). Journal of Cardiovascular Pharmacology. 57 (5): 559–67.