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ITEM PROTEIN ANTIBODY ASSAY KIT CELL
ELISA Kit for Insulin (INS)
Organism species Homo sapiens (Human)
Product No.SEA448Hu
Sample typeSerum, plasma, cell culture supernates and other biological fluids.
Format96-well strip plate
Assay length4.5 hours
Detection range39.063-2500pg/mL The standard curve concentrations used for the ELISA’s were 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.125pg/mL, 39.063pg/mL
SensitivityThe minimum detectable dose of this kit is typically less than 14pg/mL.
Specificity
This assay has high sensitivity and excellent specificity for detection of Insulin (INS).
No significant cross-reactivity or interference between Insulin (INS) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Insulin (INS) and the recovery rates were calculated by comparing the measured value to the expected amount of Insulin (INS) in samples.
Matrix Recovery range (%) Average(%)
serum(n=5)80-9680
EDTA plasma(n=5)91-9895
heparin plasma(n=5)92-10197
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Insulin (INS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Insulin (INS) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Insulin (INS) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample 1:2 1:4 1:8 1:16
serum(n=5)93-101%83-90%92-105%83-91%
EDTA plasma(n=5)86-99%99-105%80-96%92-101%
heparin plasma(n=5)98-105%96-103%82-91%93-101%
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1
Test principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Insulin (INS). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Insulin (INS). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Insulin (INS), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Insulin (INS) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Package and Components
Assay procedure summary
1. Prepare all reagents, samples and
    standards;
2. Add 100µL standard or sample to each
    well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared
    Detection Reagent A. Incubate 1 hour at
    37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent
    B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15
    -25 minutes at 37oC;
8. Add 50µL Stop Solution. Read at 450nm
    immediately.
Reagent Preparation
Results demonstration
Typical Standard Curve
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MagazineReference
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Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications
MAA448Hu22Monoclonal Antibody to Insulin (INS)WB, ICC, IHC-P, IHC-F, ELISA
NPA448Hu01Native Insulin (INS)SDS-PAGE; WB; ELISA; IP.
PAA448Hu01Polyclonal Antibody to Insulin (INS)WB, ICC, IHC-P, IHC-F, ELISA
PAA448Hu71Biotin-Linked Antibody to Insulin (INS)IHC;WB;ELISA
RPA448Hu01Recombinant Insulin (INS)SDS-PAGE; WB; ELISA; IP.
SCA448HuCLIA Kit for Insulin (INS)Chemiluminescent immunoassay
SEA448HuELISA Kit for Insulin (INS)Enzyme-linked immunosorbent assay