胰岛素(INS)检测试剂盒

胰岛素(INS)检测试剂盒

适用生物     Mus musculus (Mouse，小鼠)    

胰岛素(INS)检测试剂盒   

检测范围     123.46-10000pg/mL     灵敏度     45.5pg/mL    

样本类型     Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids    

实验时长     2.5h     实验方法     竞争抑制法    胰岛素(INS)检测试剂盒

规格     96T    

ELISA Kit for Insulin (INS)

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!

Organism species     Mus musculus (Mouse)    

胰岛素(INS)检测试剂盒

Sample type     Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids    

Format     96-well strip plate    

Assay length     2.5 hours    

Detection range     123.46-10000pg/mL The standard curve concentrations used for the ELISA’s were 10000pg/mL, 3333.33pg/mL, 1111.11pg/mL, 370.37pg/mL, 123.46pg/mL    

Sensitivity     The minimum detectable dose of this kit is typically less than 45.5pg/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)     96-103     101    

EDTA plasma(n=5)     89-97     94    

heparin plasma(n=5)     99-105     102    

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)     94-102%     96-103%     99-105%     92-101%    

EDTA plasma(n=5)     78-95%     82-97%     81-92%     85-97%    

heparin plasma(n=5)     93-102%     78-88%     96-103%     78-102%    

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     Assay Diluent A     1×12mL    

Detection Reagent B     1×120μL     Assay Diluent B     1×12mL    

Reagent Diluent     1×300μL     Stop Solution     1×6mL    

TMB Substrate     1×9mL     Instruction manual     1    

Wash Buffer (30 × concentrate)     1×20mL    

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50μL standard or sample to each well.
    And then add 50μL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37oC;
3. Aspirate and wash 3 times;
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50μL Stop Solution. Read at 450 nm immediately.

Test principle

This assay employs the competitive胰岛素(INS)检测试剂盒 inhibition enzyme immunoassay technique. A monoclonal antibody specific to Insulin (INS) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Insulin (INS) and unlabeled Insulin (INS) (Standards or samples) with the pre-coated antibody specific to Insulin (INS). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Insulin (INS) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the 胰岛素(INS)检测试剂盒concentration of Insulin (INS) in the sample.