Typical Testing Data/Standard Curve (for reference only)
Human Plasma Renin Activity ELISA Kit | PRA elisa kit
Plasma Renin Activity (PRA) ELISA
Reactivity
Human
Synonyms
Plasma Renin Activity; Plasma Renin Activity (PRA) ELISA; PRA elisa kit
Reactivity
Human
Samples
Human plasma by an enzyme immunoassay
Assay Type
Competitive
Preparation and Storage
2 - 8 degree C
Typical Testing Data/Standard Curve (for reference only)
Typical Testing Data/Standard Curve (for reference only)
Related Product Information for PRA elisa kit
Intended Uses: For the determination of Plasma Renin Activity (PRA) in human plasma by an enzyme immunoassay. For research use only,
Principle of the Assay: This kit measures PRA and the results are expressed in terms of mass of angiotensin-I (Ang-I) generated per volume of human plasma in unit time (ng/mL.h). The blood sample is collected in a tube that contains EDTA. The plasma is separated and either stored frozen or kept at room temperature for immediate use, samples should not be chilled on ice or stored at temperatures between 0 and 10 degree C during collection or processing before adjustment of pH, this could lead to overestimation of renin activity. Before the start of immunoassay a protease inhibitor and the Generation buffer is added to the plasma sample, which will prevent Angiotensin-I (Ang-I) in plasma from degradation. The pH of the plasma sample should be around 6.0 after the addition of the supplied Generation buffer. The plasma sample is split in two and the fractions are incubated at 0-4 degree C (in ice bath) and 37 degree C respectively for 90 minutes or longer, to allow the generation of Ang-I by plasma renin at 37 degree C. Optionally, the pH can be adjusted to 6.5 or 7.4. Adjustment of pH is a critical step during the assay, acidification of plasma to pH 3.3 or lower for prolonged time with subsequent return to neutral pH causes irreversible activation of the renin (Derkx et al., 1987), on the other side incubation at pH higher than 8.0 can destroy renin. During the immunoassay incubation, another set of protease inhibitors are involved, which function to stop the new generation as well as degradation of Ang-I to smaller peptides. The immunoassay of Ang-I is a competitive assay that uses two incubations, with a total assay incubation time of less than two hours. During the first incubation unlabelled Ang-I (present in the standards, controls and plasma samples) competes with biotinylated Ang-I to bind to the anti-Ang-I antibody. In the second incubation the labelled Streptavidin-HRP conjugate, binds to the immobilized Ang-I-Biotin. The washing and decanting procedures remove unbound materials. The colorimetric HRP substrate is added and, after stopping the color development reaction, the light absorbance (OD) is measured in a microwell plate reader. The absorbance values are inversely proportional to the concentration of Ang-I in the sample. A set of calibrators is used to plot a standard curve from which the concentrations of Ang-I in the samples and controls can be directly read.
Background/Introduction: Measurement of PRA is important for the research of hypertensive subjects. In particular, determination of plasma renin activity can be useful in the study of primary hyperaldosteronism (5-13% of hypertensive cases) and assist in the therapy and management of other forms of hypertension. PRA, in contrast to the determination of renin concentration, is a more accurate indicator of primary hyperaldosteronism (PHA), because of several reasons: 1. PRA is the expression of the rate of Ang-I formation through the enzymatic action of renin on its substrate, angiotensinogen, therefore PRA depends not only on renin concentration but also on the concentration of angiotensinogen which is ignored in the renin concentration assay; 2. Plasma renin concentration assay does not ensure sensitivity in low renin states, while the sensitivity of the PRA assay can be enhanced by increasing the incubation time during the generation step (Sealey et al., 2005), 3. When an inhibitor is bound to the renin active site PRA is inhibited, whereas the presence of the inhibitor does not affect the recognition of renin by currently available immunoassays, therefore total renin concentration does not always correlate with plasma renin activity (Campbell et al., 2009). Renin liberates angiotensin-I from angiotensinogen. Angiotensin-I is transformed to angiotensin-II largely in pulmonary circulation by angiotensin converting enzyme (ACE). Angiotensin-II raises blood pressure by direct arteriolar vasoconstriction, promoting sodium retention, and stimulating the secretion of aldosterone from the adrenal cortex. Aldosterone also exerts an effect to restore sodium balance and lift arterial pressure. Accurate determination of the concentration of circulating angiotensin-II is challenging because of its instability in blood samples.
Principle of the Assay: This kit measures PRA and the results are expressed in terms of mass of angiotensin-I (Ang-I) generated per volume of human plasma in unit time (ng/mL.h). The blood sample is collected in a tube that contains EDTA. The plasma is separated and either stored frozen or kept at room temperature for immediate use, samples should not be chilled on ice or stored at temperatures between 0 and 10 degree C during collection or processing before adjustment of pH, this could lead to overestimation of renin activity. Before the start of immunoassay a protease inhibitor and the Generation buffer is added to the plasma sample, which will prevent Angiotensin-I (Ang-I) in plasma from degradation. The pH of the plasma sample should be around 6.0 after the addition of the supplied Generation buffer. The plasma sample is split in two and the fractions are incubated at 0-4 degree C (in ice bath) and 37 degree C respectively for 90 minutes or longer, to allow the generation of Ang-I by plasma renin at 37 degree C. Optionally, the pH can be adjusted to 6.5 or 7.4. Adjustment of pH is a critical step during the assay, acidification of plasma to pH 3.3 or lower for prolonged time with subsequent return to neutral pH causes irreversible activation of the renin (Derkx et al., 1987), on the other side incubation at pH higher than 8.0 can destroy renin. During the immunoassay incubation, another set of protease inhibitors are involved, which function to stop the new generation as well as degradation of Ang-I to smaller peptides. The immunoassay of Ang-I is a competitive assay that uses two incubations, with a total assay incubation time of less than two hours. During the first incubation unlabelled Ang-I (present in the standards, controls and plasma samples) competes with biotinylated Ang-I to bind to the anti-Ang-I antibody. In the second incubation the labelled Streptavidin-HRP conjugate, binds to the immobilized Ang-I-Biotin. The washing and decanting procedures remove unbound materials. The colorimetric HRP substrate is added and, after stopping the color development reaction, the light absorbance (OD) is measured in a microwell plate reader. The absorbance values are inversely proportional to the concentration of Ang-I in the sample. A set of calibrators is used to plot a standard curve from which the concentrations of Ang-I in the samples and controls can be directly read.
Background/Introduction: Measurement of PRA is important for the research of hypertensive subjects. In particular, determination of plasma renin activity can be useful in the study of primary hyperaldosteronism (5-13% of hypertensive cases) and assist in the therapy and management of other forms of hypertension. PRA, in contrast to the determination of renin concentration, is a more accurate indicator of primary hyperaldosteronism (PHA), because of several reasons: 1. PRA is the expression of the rate of Ang-I formation through the enzymatic action of renin on its substrate, angiotensinogen, therefore PRA depends not only on renin concentration but also on the concentration of angiotensinogen which is ignored in the renin concentration assay; 2. Plasma renin concentration assay does not ensure sensitivity in low renin states, while the sensitivity of the PRA assay can be enhanced by increasing the incubation time during the generation step (Sealey et al., 2005), 3. When an inhibitor is bound to the renin active site PRA is inhibited, whereas the presence of the inhibitor does not affect the recognition of renin by currently available immunoassays, therefore total renin concentration does not always correlate with plasma renin activity (Campbell et al., 2009). Renin liberates angiotensin-I from angiotensinogen. Angiotensin-I is transformed to angiotensin-II largely in pulmonary circulation by angiotensin converting enzyme (ACE). Angiotensin-II raises blood pressure by direct arteriolar vasoconstriction, promoting sodium retention, and stimulating the secretion of aldosterone from the adrenal cortex. Aldosterone also exerts an effect to restore sodium balance and lift arterial pressure. Accurate determination of the concentration of circulating angiotensin-II is challenging because of its instability in blood samples.
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Product Notes
The Human PRA (Catalog #AAA495044) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA495044 ELISA Kit recognizes Human PRA. It is sometimes possible for the material contained within the vial of "Plasma Renin Activity, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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