Rat Insulin Degrading Enzyme, IDE ELISA Kit

Catalog No.: BZEK2424-48

Size: 48T

Range: 10 ng/L - 160 ng/L

Sensitivity: 1 ng/L

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of IDE in rat serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Insulin-degrading enzyme, also known as IDE, is an enzyme. Known alternatively as insulysin or insulin protease, IDE is a large zinc-binding protease of the M16 metalloprotease family known to cleave multiple short polypeptides that vary considerably in sequence. Other members of this family include the mitochondrial processing peptidase and presequence protease. The gene IDE encodes protein Insulin-degrading enzyme. The human gene IDE has 28 exons and is located at chromosome band 10q23-q25.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- IDE antibody was pre-coated onto 48-well plates. And the HRP conjugated anti- IDE antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) 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 IDE amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of IDE can be calculated.

Kit components

1. One 48-well plate pre-coated with anti- rat IDE antibody

2. Standard: 0.5ml (320 ng/L)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (20×): 20 ml. Dilution: 1:20

5. Sample diluent buffer: 3 ml

6. HRP conjugated anti- rat IDE antibody (RTU): 3ml

7. Stop solution: 3 ml

8. TMB substrate A:  3ml

9. TMB substrate B:  3ml

10. Plate sealer:   2

11. Hermetic bag:  1

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. Absorbent filter papers
8. Plastic or glass container with volume of above 1L