Mouse Cystatin C ELISA Kit

Catalog No.: BEK1040

Size: 96T

Range: 312 pg/ml-20,000 pg/ml

Sensitivity < 10 pg/ml

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

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

Introduction

Cystatin C or cystatin 3, a protein encoded by the CST3 gene, is mainly used as a biomarker of kidney function. It is a single 120-residue polypeptide with a molecular mass of approximately 13.26 kD. Cystatin C plays a role in network reorganization in the epileptic dentate gyrus, especially in granule cell dispersion and guidance of migrating newborn granule cells. It is an alternative measure of kidney function that may have prognostic importance among elderly persons who do not meet standard criteria for chronic kidney disease. Among elderly persons without chronic kidney disease, cystatin C is a prognostic biomarker of risk for death, cardiovascular disease, and chronic kidney disease, thereby signaling a state of 'preclinical' kidney dysfunction.

Principle of the Assay 

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

Kit components

1. One 96-well plate pre-coated with anti-Mouse Cystatin C antibody
2. Lyophilized Mouse Cystatin C standards: 2 tubes (20ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse Cystatin C 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