Human Insulin Pincer^TM Assay

Catalog No: IH1010

This human insulin Pincer^TM assay kit is to be used for the measurement of insulin levels in biological buffers and culture medium, for example, to monitor the insulin production in the cultured or extract of islets of Langerhans.

Insulin is a hormone responsible for regulating the metabolism of glucose and most amino acids in human body. It is synthesized in the β-cells of the islets of Langerhans, and is released after removal of the C-peptide from proinsulin¹. Human insulin consists of a 21-residue A-chain and a 30-residue B-chain, linked by two disulfide bonds. The insulin hexamer forms in the presence of Zn²⁺ ions, which is the inactive form with long-term stability^2,3. Insulin plays important roles on human metabolism by controling cellular intake of glucose, increasing DNA replication and protein synthesis, regulating enzyme activities. Diabetes mellitus develops when the body does not produce enough (type I) or properly respond to (type II) insulin.

This assay consists two human insulin specific antibodies which recognize different epitopes on human insulin. Each antibody is modified with short complementary single-stranded DNAs which associate to form a duplex in the presence of the human insulin. 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 insulin concentration present in the samples⁴. This assay is simple (one step) and fast (30 min).

Human Insulin Pincer^TM Assay - Standard Curves

The following human insulin standard curve was prepared using the Human Insulin Pincer^TM 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 Insulin Pincer^TM Assay – Sensitivity And Specificity

Sensitivity: The minimum detectable dose of human insulin using a standard curve is 0.5 ng/mL.
Specificity: No cross reaction observed.
Detection Range: 0.5 ng/ml – 40 ng/ml.

Reference:

1. Steiner DF, Oyer PE (1967). The biosynthesis of insulin and a probable precursor of insulin 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 insulin hexamer, Biochemistry, 36 (31): 9409–22.
3. Dunn MF, (2005) Zinc-ligand interactions modulate assembly and stability of the insulin hexamer - a review, BioMetals, 18 (4): 295–303.
4. Heyduk T, & Heyduk, E, (2005). Nucleic acid-based fluorescence sensors for detecting proteins. Anal. Chem., 77:1147-1156.
5. Heyduk, E., Moxley, MM., Salvatori, A., Corbett, JA. and Heyduk, T.Homogeneous insulin and C-peptide sensors for rapid assessment of insulin and C-peptide secretion by the islets. Diabetes. 2010 Jul 9.

RESEARCH USE:

For research use only, not for use in diagnostic procedures.

Patent pending.

For 96-well format please inquire. Click Here.

Product name	Measurements	Cat. #	Protocol	MSDS	Price	Order
Human Insulin PincerTMAssay kit	100	IH1010-100			$180
Human Insulin PincerTMAssay kit	384	IH1010-384			$320
Human Insulin PincerTMAssay kit	384 x 2	IH1010-384 x 2			$580
Human Insulin PincerTMAssay kit	384 x 3	IH1010-384 x 3			$800
384-well round-bottom low volume non-binding surface black polystyrene microplate	1 plate	163301	--		$9.50