Mercodia Rat Insulin ELISA
10-1250-01 / 10-1250-10
www.mercodia.com
Intended use
A high quality enzyme immunoassay for the quantification of rat insulin in serum or plasma.
Test principle
Mercodia Rat Insulin ELISA is a solid phase two-site enzyme immunoassay based on the sandwic technique, in which two monoclonal antibodies are directed against separate antigenic determinants on the insulin molecule. Insulin in the sample reacts with anti-insulin antibodies bound to microtitration wells and peroxidase-conjugated anti-insulin antibodies in the solution.
Test characteristics
Sensitivity
The detection limit is ≤ 0.15 μg/L as determined with the methodology described in ISO11843-Part 4.
Recovery
Recovery upon addition is 80 - 93 % （86 %）.
Recovery upon dilution is 83 - 111 % （96 %）.
Precision
Each sample was analyzed in 4 replicates on 16 different
occasions.
Performance limitations
Grossly lipemic, icteric or haemolyzed samples do not interfere in the assay.
Specificity
The following cross-reactions have been tested:
Human Insulin 167 %
Human proinsulin 75 %
Human C-peptide < 0.05 %
Insulin lispro 167 %
IGF-I < 0.02 %
IGF-II < 0.02 %
Rat C-peptide < 0.001 %
Rat proinsulin 7 %
Porcine insulin 628 %
Ovine insulin 256 %
Bovine insulin 110 %
Samples
Serum, EDTA and heparin plasma can be used.
Measurement range
0.15 - 5.5 μg/L
Catalog No
10-1250-01 1 x 96 wells
10-1250-10 10 x 96 wells
Sample
Mean value
Coefficient of variation
（μg/L）
within assay %
between assay %
total assay %
1
2
3
0.47
0.96
2.8
5.1
3.1
2.8
10
4.4
3.2
6.1
3.5
5.3
Summary of protocol
? Add 10 μL Calibrators, controls and samples
? Add 100 enzyme conjugate solution
? Incubate 2 hours on shaker at room temperature
? Wash plate 6 times
? Add 200 Substrate TMB
? Incubate 15 minutes at room temperature
? Add 50 μL Stop Solution
Shake for approximay 5 seconds on shaker
? Measure at 450 nm
Global Headquarters
Mercodia AB, Sweden
Phone +46 18 57 00 70
info@mercodia.com
US Sales Office
Mercodia Inc
Phone （336） 725-8623
info_usa@mercodia.com
2010-05-19, Version 1, Article No 32-4167

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