Prouroguanylin Human ELISA

Research topic

Others, Renal disease

Features

• The total assay time is less than three hours.
• The kit measures total serum prouroguanylin.
• Quality controls are human serum based. No animal sera are used.

Storage/Shipping

Store the kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Summary

Prouroguanylin (about 9, 7 kDa) is a biologically inactive form of uroguanylin circulating in a bloodstream. Uroguanylin is a small –molecular-weight peptide which has been shown to participate in the regulation of salt and water homeostasis in mammals via cGMP-mediated processes in the intestine, kidney and other epithelia. Prouroguanylin levels are markedly increased in chronic renal failure. Uroguanylin/prou­roguanylin levels also increased in the nephrotic syndrome. Studies of pathogenesis of colorectal cancer demonstrate that prouroguanylin may serve as marker of colon tumors in the body. Recent experiments also refer to possibility of prouroguanylin to play a significant role at diagnostic and treatment of heart diseases.

Assay format

Sandwich ELISA, Biotin-labelled antibody

Sample requirements

20 µl/well

Applications

Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum

Calibration Curve

Calibration range

0.6 – 20 ng/ml

Limit of detection

Analytical Limit of Detection is calculated from the real prouroguanylin values in wells and is 0.13 ng/ml.

Limit of quantification

Assay Sensitivity takes the dilution of samples into consideration and is calculated according to the formula: Assay Sensitivity = Analytical Limit of Detection x sample dilution = 0.13 ng/ml x 5 = 0.65 ng/ml

Intra-assay (Within-Run, n=8)

CV = 3.85 %

Inter-assay (Run-to-Run, n=8)

CV = 3.50 %

Spiking Recovery

101.4 %

Dilution Linearity

104.5 %

Cross-Reactivity

Human, Monkey

References to summary

• Fan X. Wang Y, London RM, Eber SL, Krause WJ, Freeman RH, Forte LR: Signaling Pathways for Guanylin and Uroguanyiln in the Digestive, Renal, Central Nervous, Reproductive, and Lymphoid Systems. Endocrinology 138, 4636–4648 (1997).
• Forte LR: A novel role for uroguanylin in the regulation of sodium balance. Am Clin Invest 112, 1138–1141 (2003).
• Forte LR, Freeman RH, Krause WJ, London RM: Guanylin peptides: cyclic GMP signalling mechanisms. Braz J med Biol Res 32(11), 1329–1336 (1999).
• Forte LR, London RM, Freeman RH, Krause WJ: Guanylin peptides: renal actions mediated by cyclic GMP. Am J Physiol Renal Physiol 278, 180–191 (2000).
• Hamra FK, Fan X, Krause WJ, Freeman RH, Chin DT, Smith ChE, Currie MG, Forte LR: Prouroguanylin and proguanylin: Purification from Colon, Structure, and Modulation of Bioactivity by Proteases. Endocrinology 137, 257–265 (1996).
• Hidaka Y, Shimono Ch, Ohno M, Okumura N, Adermann K, Forshmann W-G, Shimonishi Y: Dual Function of the Propeptide of Prouroguanylin in the Folding of the Mature Peptide. J Biol Chem 257, 25155–25162 (2000).
• Shailubhai K, Yu HH, Karunanandaa K, Wang JY, Eber SL, Wang Y, Joo NS, Kim HD, Miedema BW, Abbas SZ, Boddupalli SS, Currie MG, Forte LR: Uroguanylin Treatment Suppresses Polyp Formation in the APCMin/+ Mouse and Induces Apoptosis in Human Colon Adenocarcinom Cells via Cyclic GMP. Cancer research 60, 5151–5157 (2000).