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Human C-peptide PincerTM Assay |
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Catalog No: CP1013 This human C-peptide PincerTM assay kit is to be used for the measurement of insulin levels in biological buffers. Human C-peptide is a 31 amino acids peptide with a molecular mass of approximately 3000 daltons. C-peptide plays an important role in insulin biosynthesis by facilitating the correct folding of the proinsulin molecule and linking the a-chain and b-chain together in proinsulin1,2. C-peptide exhibits several cellular functions including stimulation of Na+, K+-ATPase3, increment of NO production4, activation of mitogen activated protein kinases (MAPKs)5 and PI-3-kinase6,7, and enhancement of expression and translocation of nuclear factor-κB (NF-κB)7. Monitoring c-peptide level provides a better way to quantify insulin secretion since c-peptide and insulin are secreted in equimolar amounts, and c-peptide has longer half-life than insulin. This assay consists two human c-peptide specific antibodies which recognize different epitopes on human c-peptide. Each antibody is modified with short complementary single-stranded DNAs which associate to form a duplex in the presence of the human c-peptide. FRET (Fluorescence resonance energy transfer) is initiated between the two fluorophores added to the end of the two DNAs. The amount of FRET signal is associated with the human c-peptide concentration present in the samples8. This assay is simple (one step) and fast (30 min).
Human C-peptide PincerTM Assay - Standard Curves The following human C-peptide standard curve was prepared using the Human C-peptide PincerTM Assay kit. This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. Each user should obtain his own data and standard curve.
Human C-peptide PincerTM Assay – Sensitivity And Specificity Sensitivity: The minimum detectable dose of human c-peptide using a standard curve is 2 ng/mL.
Reference: 1.Steiner DF, Oyer PE, (1967). The biosynthesis of c-peptide and a probable precursor of c-peptide by a human islet cell adenoma. Proc. Natl. Acad. Sci. U.S.A. 57 (2): 473–480 2. Chang X, Jorgensen AM, Bardrum P, Led JJ, (1997). Solution structures of the R6 human c-peptide hexamer. Biochemistry, 36 (31): 9409–22. 3. Ohtomo Y, Aperia A, Sahlgren B, et al., (1996). C-peptide stimulates rat renal tubular Na+,K+-ATPase activity in synergism with neuropeptide Y. Diabetologia. 39:199–205. 4. Kitamura T, Kimura K, Makondo K, et al., (2003). Proinsulin C-peptide increases nitric oxide production by enhancing mitogen-activated protein-kinasedependent transcription of endothelial nitric oxide synthase in aortic endothelial cells of Wistar rats. Diabetologia. 46:1698–1705. 5. T. Kitamura, K. Kimura, B. D. Jung, et al., (2001). Proinsulin Cpeptide rapidly stimulates mitogen activated protein kinases in Swiss 3T3 fibroblasts: requirement of protein phosphoinositide 3-kinase and pertussi toxin-sensitive G protein. Biochemical Journal, 355:123–129. 6. N. M. Al-Rasheed, F.Meakin, E. L. Royal, et al., (2004). Potent activation of multiple signalling pathways by C-peptide in opossum kidney proximal tubular. Diabetologia, 47(6): 987– 997. 7. Z.-G. Li, W. Zhang, and A. A. F. Sima, (2003). C-peptide enhances insulin-mediated cell growth and protection against high glucose-induced apoptosis in SH-SY5Y cells. Diabetes/ Metabolism Research and Reviews, 19(5): 375–385. 8. Heyduk T, & Heyduk, E, (2005). Nucleic acid-based fluorescence sensors for detecting proteins. Anal. Chem., 77:1147-1156.
RESEARCH USE: For research use only, not for use in diagnostic procedures. Patent pending. For 96-well format please inquire. Click Here.
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