ELISA by Species

Product#: ELISA _by _Species
ELISA _by _Species
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* Either 96 OR 48 Samples 
* Cover Major 14 Different Vertebrates 
* Pan-species ELISA Available
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* Targeting Proteins, Ligands, Antibodies & Phosphorylation
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Human Proinsulin (PI) ELISA Kit
two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.
Item Standard Test

The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of PI in human serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.

Identification Colorimetric Positive
Composition Traditional ELISA Kit Ready-to-Use ELISA KIT Conform
Pre-coated, ready to use 96-well strip plate 1 Pre-coated, ready to use 96-well strip plate 1
Plate sealer for 96 wells 2 Plate sealer for 96 wells 2
Standard 2 Standard 2
Diluents buffer 1×45mL Standard Diluent 1×20mL
Detection Reagent A 1×120μL Detection Solution A 1×12mL
Detection Reagent B 1×120μL Detection Solution B 1×12mL
TMB Substrate 1×9mL TMB Substrate 1×9mL
Stop Solution 1×6mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1 Instruction manual 1
Test principle

The microtiter plate provided in this kit has been pre-coated with an antibody specific to PI. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to PI. 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 PI, 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 PI in the samples is then determined by comparing the O.D. of the samples to the standard curve.


Matrices listed below were spiked with certain level of recombinant PI and the recovery rates were calculated by comparing the measured value to the expected amount of PI in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-102 91
EDTA plasma(n=5) 81-98 89
heparin plasma(n=5) 80-89 84

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of PI 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) 82-96% 83-98% 81-99% 93-101%
EDTA plasma(n=5) 88-101% 86-95% 90-102% 80-93%
heparin plasma(n=5) 80-91% 82-90% 95-104% 79-95%

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level PI were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level PI were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%


The stability of ELISA 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 conditions.
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

Assay procedure summary

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

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