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ITEM PROTEIN ANTIBODY ASSAY KIT CELL
CLIA Kit for Insulin (INS)
Organism species Homo sapiens (Human)
Product No.SCA448Hu
Sample typeSerum, plasma and other biological fluids
Format96-well strip plate
Assay length4.5 hours
Detection range3.429-2500pg/mL The standard curve concentrations used for the ELISA’s were 2500pg/mL, 833.33pg/mL, 277.78pg/mL, 92.59pg/mL, 30.864pg/mL, 10.288pg/mL, 3.429pg/mL
SensitivityThe minimum detectable dose of this kit is typically less than 1.56pg/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)96-103101
EDTA plasma(n=5)91-9895
heparin plasma(n=5)78-10498
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)78-98%78-92%85-95%87-101%
EDTA plasma(n=5)82-96%80-89%95-105%87-94%
heparin plasma(n=5)91-99%90-97%80-95%99-105%
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
Substrate A 1×10mL Substrate B 1×2mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1
Test principle
The microtiter plate provided in this kit has been pre-coated with an antigen. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated secondary antibody. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Insulin (INS) level in the sample or standard.
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 100µL Substrate Solution. Incubate     10 minutes at 37oC;
8. Read RLU value immediately.
Reagent Preparation
Results demonstration
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Benchmark price
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications
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