Insulin High Range Assay kit
Insulin quantification for higly concentrated samples
Insulin Mouse Serum Assay kit HTRF®
The Insulin Mouse Serum Assay kit allows for a reliable time-saving quantification of insulin in serum and plasma samples from mice. A perfect tool in diabetes research.
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Faster and more convenient than ELISA
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Low sample consumption
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No-wash
Overview
For years, researchers have sought the ideal insulin assay that would combine absolute specificity with sensitivity, accuracy, low sample consumption, and ease-of-use. The search has come to an end! This serum insulin assay offers superior specificity and sensitivity for a reliable quantification of insulin in serum and plasma samples. Built on a truly homogeneous protocol, the assay does not require any washing or complex extraction steps, leading to significant time savings over ELISA methods.
Benefits
- HIGH SENSITIVITY
- HIGH SPECIFICITY
- DIABETES RESEARCH
Assay principle
Insulin is measured using a sandwich immunoassay of two monoclonal antibodies, one labelled with Cryptate (donor) and the other with XL665 (acceptor). The intensity of the FRET signal obtained is proportional to the concentration of insulin in the sample.
Assay Protocol
10 µL of cell supernatants are supplemented with 10 µL of acceptor-labelled antibodies and 5 µL of donor-labelled antibodies (or 15 µL of pre-mixed reagents for a single dispensing step). The mix is then incubated from 4h to overnight at RT and results are read on an HTRF compatible reader.
Sensitivity
The detection limit* is calculated as two standard deviations above the Calibrator 0. The HTRF® Insulin assay was calibrated against the NIBSC international standard 66/304. The following equivalence was found: 1 ng Insulin HTRF® represent 1 ng Insulin NIBSC 66/304 (i.e.: 2.3x10-05 IU). Samples were incubated overnight at room temperature. The assay ranges from 0,24 to 8 ng/mL and the detection limit measured was 0,064 ng/mL.
Precision intra-assay
| Mouse serum sample | Mean [Insulin] (ng/mL) | CV |
| 1 | 0.25 | 7.6% |
| 2 | 0.73 | 3.7% |
| 3 | 0.9 | 5.4% |
| 4 | 2.84 | 4.6% |
| 5 | 5.17 | 4.0% |
| Mean CV | 5.1% |
Precision inter-assay
| Mouse serum sample | Mean [Insulin] (ng/mL) | CV |
| 1 | 0.37 | 14.6% |
| 2 | 0.67 | 9.5% |
| 3 | 0.9 | 4.0% |
| 4 | 1.1 | 8.1% |
| 5 | 6.9 | 9.2% |
| Mean CV | 9.1% |
Dilutional linearity
| Mouse serum sample | Dilution factor | [Insulin] expected (ng/mL) | [Insulin] measured (ng/mL) | Recovery |
| 1 | 1 | - | 0.97 | - |
| 2.5 | 0.388 | 0.368 | 95% | |
| 5 | 0.194 | 0.188 | 97% | |
| 2 | 1 | - | 4 | - |
| 2.5 | 1.6 | 1.57 | 98% | |
| 5 | 0.8 | 0.9 | 110% | |
| 3 | 1 | - | 7.1 | - |
| 2.5 | 2.84 | 3 | 105% | |
| 5 | 1.42 | 1.66 | 117% | |
| Mean Recovery | 103.7% |
Spike and recovery
| Mouse serum sample | [Insulin] added (ng/mL) | [Insulin] expected (ng/mL) | [Insulin] measured (ng/mL) | Recovery |
| 1 | 0 | - | 0.75 | - |
| 2.3 | 3.05 | 3.6 | 118% | |
| 4.8 | 5.55 | 6.4 | 115% | |
| 2 | 0 | - | 2.5 | - |
| 2.4 | 4.9 | 4.8 | 98% | |
| 4.8 | 7.3 | 8 | 110% | |
| 3 | 0 | - | 4.2 | - |
| 0.8 | 5 | 5.4 | 108% | |
| 1.6 | 5.8 | 6.2 | 107% | |
| Mean Recovery | 109.3% |
Correlation with leading ELISA
The HTRF Mouse serum insulin assay showed an excellent correlation to the leading absorbance and chemiluminescent ELISA, both on sera with low titers and high titers of insulin. With a slope of 1.1 and 0.9 respectively, the regression line was close to the ideal line, demonstrating the assay reliability.
Validation on normal mouse serum
C57BL/6J mice were randomized into two groups of 7 animals. One group was fasted overnight while the second one was normally fed. The mice were sacrificed and the blood was collected for serum preparation. The insulin concentration was determined in each sample using the HTRF Mouse Serum Insulin kit reagents.
Validation on diabetic mouse plasma
A group of five db/db diabetic male mice and a group of five db/+ control male mice were fasted overnight. The mice were sacrificed and the blood was collected through cardiac puncture for K3 EDTA-plasma preparation. The insulin concentration of each sample was determined using the HTRF Mouse Serum Insulin kit reagents.
Validation on obese mouse serum
A group of eight ob/ob obese mice and a group of eight ob/+ control mice were sacrificed and the blood was collected through cardiac puncture for serum preparation. The insulin concentration of each sample was determined using the HTRF Mouse Serum Insulin kit reagents.
In collaboration with Elixir - Scientific Presentations
Designed for in vivo experiments on mouse models secreting all insulin levels. - Flyers
The next level in Insulin quantification assays
Compare easily the 3 Cisbio kits for insulin quantification - Guides
Product Insert Insulin (m)serum Kit / 62IN3PEF-62IN3PEB
62IN3PEF-62IN3PEB - Product Insert
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
A guide to help you in your diabetes projects
A comprehensive listing of Type II diabetes pathways - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
Safety Data Sheet (DEU) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (ELL) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (FRA-FR) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (ITA) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (SPA) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (ENG-GB) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (ENG-US) Insulin (m)serum Kit / 62IN3PEB
62IN3PEB - Safety Data Sheet
Safety Data Sheet (DEU) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (ELL) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (FRA-FR) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (ITA) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (SPA) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (ENG-GB) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Safety Data Sheet (ENG-US) Insulin (m)serum Kit / 62IN3PEF
62IN3PEF - Safety Data Sheet
Plate Reader Requirement
Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.
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