Rat Insulin ELISA Kit

Catalog No.: BEK1324

Size: 96T

Range: 1.56 µIU/ml - 100 µIU/ml

Sensitivity < 2 µIU/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months. 

Application: For quantitative detection of Insulin in rat serum, body fluids, tissue lysates or cell culture supernatants.

Introduction

Insulin, synthesized by the beta cells of the islets of Langerhans, consists of 2 dissimilar polypeptide chains, A and B, which are linked by 2 disulfide bonds. The human insulin gene contains 3 exons; exon 2 encodes the signal peptide, the B chain, and part of the C peptide, while exon 3 encodes the remainder of the C peptide and the A chain. Insulin has a potent acute antiinflammatory effect, including a reduction in intranuclear NF-kappa-B, an increase in IKB, and decreases in the generation of reactive oxygen species. It causes cells in the liver, muscle, and fat tissue to take up glucose from the blood, storing it as glycogen inside these tissues, and improved insulin-stimulated glucose uptake after endurance training results from hemodynamic adaptations as well as increased cellular protein content of individual insulin signaling components and molecules involved in glucose transport and metabolism.

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- Insulin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- Insulin polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Insulin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Insulin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-rat Insulin antibody
2. Lyophilized rat Insulin standards: 2 tubes (4000 µIU / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-rat Insulin antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L