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Insulin assay

The HTRF® Insulin assay is an 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.

Features

  • Cost-effective alternative to ELISA
  • Homogeneous assay
  • Assay results in under 3 hours
  • 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
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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 tests62INSPEB
Insulin kit - 20,000 tests62INSPEC

Companion products

DescriptionCat. noProduct insertMSDS
Insulin calibrator 62INSCDA
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Insulin control 62INSTDA
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