Mouse Resistin ELISA Kit

Catalog No.: BEK1197

Size: 96T

Range: 31.2 pg/ml-2000 pg/ml

Sensitivity < 2 pg/ml

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

Application: For quantitative detection of Resistin in mouse serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Resistin also known as adipose tissue-specific secretory factor or C/EBP-epsilon-regulated myeloid-specific secreted cysteine-rich protein (XCP1) is a cysteine-rich protein that in humans is encoded by the RETN gene. The gene comprises 4 exons, the first of which is untranslated, and spans approximately 1,750 bp. The length of the resistin pre-peptide in human is 108 amino acid residues and in the mouse and rat it is 114 aa; the molecular weight is ~12.5 kDa. Resistin is a cytokine whose physiologic role has been the subject of much controversy regarding its involvement with obesity and type II diabetes mellitus (T2DM).

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Resistin polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Resistin 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 Resistin amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Resistin can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-mouse Resistin antibody
2. Lyophilized Mouse Resistin standards: 2 tubes (10 ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse Resistin 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