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HTRF Mouse Insulin Serum Detection Kit, 200 Assay Points

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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Feature Specification
Application Protein Quantification
Sample Volume 5 µL

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.

Click to copy promo code to clipboard.
img-icon-save-10-black.svg

March lab savings - buy on Revvity.com or Revvity punchout to SAVE 10% with promo code!

MARCH10

Sale ends 3/24. Terms and conditions apply.

Product Variants
Unit Size: 200 assay points
Part #:
62IN3PEF
List Price
USD 895.21
Your online price:
Unit Size: 1,000 assay points
Part #:
62IN3PEB
List Price
USD 3,598.03
Your online price:
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption, and disposal requirements under European REACH regulations (EC 1907/2006).

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.

Specifications

Application
Protein Quantification
Brand
HTRF
Detection Modality
HTRF
Product Group
Kit
Sample Volume
5 µL
Shipping Conditions
Shipped Ambient
Target Class
Biomarkers
Target Species
Mouse
Technology
TR-FRET
Therapeutic Area
Metabolism/Diabetes
Unit Size
200 assay points

Video gallery

Citations

How it works

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.

 

1biomarkers-how-it-works-assay-principle-insulin-high-range-62in1peg-62in1peh.svg

 

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.

 

2biomarkers-how-it-works-assay-protocol-insulin-high-range-62in1peg-62in1peh.svg

 

Assay details

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.

 

3-3specifications-insulin-mouse-serum-1.svg

 

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.

4-3specifications-insulin-mouse-serum-2.svg
5-3specifications-insulin-mouse-serum-3.svg

 

Assay validation

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.

 

6-3assay-validation-insulin-mouse-serum-1.svg

 

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.

 

7-3assay-validation-insulin-mouse-serum-2.svg

 

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.

 

8-3assay-validation-insulin-mouse-serum-3.svg

 

Resources

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