ELISA Kit for Insulin (INS)

Brand

Cloud-Clone

short description

Enzyme-linked immunosorbent assay for Antigen Detection.
Research focus: Metabolic pathway; Endocrinology; Hormone metabolism; ;

spezie

Rhesus monkey (Simian)

Cross Reaction

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.

detection target

Insulin
coating antibody: Mouse

sensitivity

The minimum detectable dose of this kit is typically less than 44.9 pg/mL

detection range

123.46-10000 pg/mL

certificate

ISO 9001:2008, ISO 13485:2003

sample type

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

pack size

48T, 96T, 5x96T, 10x96T

Reproducibility

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%

Assay time

2h

storage and Stability

All the kits should stored according to the manual.
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree upon receipt while the others should be at 4 degree.
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.

Principle of the Elisa

Competitive Inhibition
Test principle: This assay employs the competitive 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 concentration of Insulin (INS) in the sample.

Kit Content

Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard: 2
Standard Diluent: 1x20mL
Detection Reagent A: 1
Assay Diluent A: 1x12mL
Detection Reagent B: 1x120µ L
Assay Diluent B: 1x12mL
Reagent Diluent: 1x300µ L
Stop Solution: 1x6mL
TMB Substrate: 1x9mL
Wash Buffer (30 x concentrate): 1x20mL
Instruction manual:1

Assay procrdure summery

1. Prepare all reagents, samples and standards;
2. Add 50µ L standard or sample to each well.
        And then add 50µ L prepared Detection Reagent A immediately.
        Shake and mix. Incubate 1 hour at 37° C;
3. Aspirate and wash 3 times;
4. Add 100µ L prepared Detection Reagent B. Incubate 30 minutes at 37° C;
5. Aspirate and wash 5 times;
6. Add 90µ L Substrate Solution. Incubate 10-20 minutes at 37° C;
7. Add 50µ L Stop Solution. Read at 450 nm immediately.