You are here : Home / Products / Biomarkers / Biomarkers / Insulin

Insulin assay

Insulin, a peptide hormone, is secreted by pancreatic ß-cells and is a key regulator in glucose homeostasis. Insulin deficiency leads to insulin-dependent (type 1) diabetes, whereas resistance to insulin action is common in non-insulin-dependent diabetes (type 2), obesity, and endocrine dysfunctions. The HTRF® Insulin assay is a homogeneous assay for rapid and accurate measurement of insulin in rat, mouse, pig and human samples. This ready-to-use kit is available with two standard ranges. The choice of the protocol will depend on the nature of the samples:

  • High Range protocol (10 to 100 ng/mL) for samples from glucose perfused pancreas
  • Sensitive Range protocol (0.2 to 10 ng/mL) for pancreatic ß-cell supernatants and serum samples

Features

  • Cost-effective alternative to ELISA
  • Homogeneous assay
  • Assay results in under 3 hours
  • Suitable for HTS
  • Excellent reproducibility: CV<4%
  • Small sample volume needed (as low as 5-10 µL)
  • Two standard ranges: High Range (10-100 ng/mL) and Sensitive Range (0.2-10 ng/mL). Amenable to automation and HTS

Applications

  • Insulin measurement
  • Screening of insulinotropic molecules
  • Diabetes assessment
ask for a quote contact us

Related applications and services

The HTRF Insulin assay can be used to measure rat, pig, mouse or human insulin in a variety of media, including cell supernatants and serum. The assay is a sandwich immunoassay that uses two monoclonal antibodies, one labeled with Eu3+-Cryptate and one labeled with XL665, that recognize distinct epitopes. The assay is performed in a single step and only requires one incubation. The two detection reagents can be premixed and pre-plated, and the plates frozen for later use. This feature is possible due to the stability and robustness of HTRF reagents, and gives a practical advantage for screening.

Detection limit: 0.20 ng/mL (Sensitive Range protocol)
4 ng/mL (High Range protocol)
Dynamic range: 0.3 to 10 ng/mL (Sensitive Range protocol)
3 to 100 ng/mL (High Range protocol)
S/B: 18 in 96- or 384-well plates
Z’: as high as 0.89
Specificity: rat, mouse, porcine & human insulin

Insulin standard curves obtained with the Sensitive Range protocol and performed using the designated final assay volumes and plate types.

Insuline standard curve

Rat and mouse models are commonly used in diabetes research. In these models, insulin secretion is monitored on pancreatic cell lines as well as in biological fluids. Cisbio Bioassays' HTRF insulin assay has been validated using a number of experimental systems, including screening of insulinotropic molecules on pancreatic β-cells under HTS conditions and monitoring of in vivo insulin and glucose production.

Pancreatic β-cells

INS-1E pancreatic β-cells underwent starvation for 48 hours prior to stimulation with glucose and additional insulinotropic agents. Insulin concentrations in cell supernatants were determined using the HTRF Insulin assay following the Sensitive Range protocol. As shown in figure 1, glucose alone stimulated measurable insulin levels. As expected, the addition of insulinotropic agents stimulated increased insulin production compared to glucose alone.

biomarker insulin performance 1

Figure 1: 104 INS-1E pancreatic β-cells /100 µL/well were plated in a 96-well tissue-culture treated plate. Cells were stimulated with 1mM glucose in combination with different compounds. 50 µL of cell supernatants were tested using standard HTRF insulin assay conditions.

In vivo Studies

In these studies rats were anesthetized and two catheters inserted, one used to obtain blood samples and the other used for the injection of glucose. Blood samples were taken at the time of injection and at various intervals thereafter. Samples were then tested for glucose and insulin levels (Figure 2). Induction of insulin was easily detected and correlated with glucose levels, using as little as 50µl blood samples.

biomarker insulin performance 2

Figue 2: Male Wistar rats were injected via catheter with glucose (3g/kg).T=0 is the time of glucose injection. Samples were dispensed into a 96 half-well plate (50 µL/well) and insulin levels measured using the standard HTRF insulin assay protocol.

 

Ordering Info

DescriptionCat. noProduct insertMSDS
Insulin kit - 1,000 tests62INSPEBpdfpdf
Insulin kit - 20,000 tests62INSPECpdfpdf

Companion products

DescriptionCat. noProduct insertMSDS
Insulin calibrator62INSCDA-pdf
Insulin control62INSTDA-pdf

Title

Related products

Download

Title

Related products

Download

  • Allosteric Modulation of Endogenous Metabolites as an Avenue for Drug Discovery
    Wootten D, Savage EE, Valant C, May LT, Sloop KW, Ficorilli J, Showalter AD, Willard FS, Christopoulos A, Sexton PM.
    Mol Pharmacol. 2012;82(2):281-90.
    Insulin assay
  • Upregulation of β-cell genes and improved function in rodent islets following chronic glucokinase activation
    Gill D, Brocklehurst KJ, Brown HW, Smith DM
    J Mol Endocrinol. 2011 Jul 18;47(1):59-67
    Insulin assay
  • Excessive Food Intake, Obesity and Inflammation Process in Zucker fa/fa Rat Pancreatic Islets
    Chentouf M, Dubois G, Jahannaut C, Castex F, Lajoix AD, Gross R, Peraldi-Roux S
    PLoS One. 2011;6(8):e22954
    Insulin assay
  • Designed inhibitors of insulin-degrading enzyme regulate the catabolism and activity of insulin.
    Leissring MA, Malito E, Hedouin S, Reinstatler L, Sahara T, Abdul-Hay SO, Choudhry S, Maharvi GM, Fauq AH, Huzarska M, May PS, Choi S, Logan TP, Turk BE, Cantley LC, Manolopoulou M, Tang WJ, Stein RL, Cuny GD, Selkoe DJ.
    PLoS One. 2010 May 7;5(5):e10504.
    Insulin assay
  • Quercetin potentiates insulin secretion and protects INS-1 pancreatic β-cells against oxidative damage via the ERK1/2 pathway
    Youl E, Bardy G, Magous R, Cros G, Sejalon F, Virsolvy A, Richard S, Quignard JF, Gross R, Petit P, Bataille D, Oiry C
    Br J Pharmacol. 2010 Oct;161(4):799-814.
    Insulin assay
  • Loss of Lkb1 in adult beta cells increases beta cell mass and enhances glucose tolerance in mice
    Fu A, Ng AC, Depatie C, Wijesekara N, He Y, Wang GS, Bardeesy N, Scott FW, Touyz RM, Wheeler MB, Screaton RA.
    Cell Metab. 2009 Oct;10(4):285-95.
    Insulin assay

    • Title

    Related products

    Download

  • A high-throughput HTRF® assay for human/rat/mouse insulin
    Claret EJ , Leroy C, Pablo2 AL, Drexler C, Tousch D, Ribes D, Gross R, Seguin P
    SBS 10th Annual Conference. September 2004, Orlando, USA
    Insulin assay
     

    • Title

    Related products

    Download

    • Title

    Related products

    Download

    • iba-worldwide.com
    © 2012 Cisbio.com - All Rights Reserved