Human Cathepsin B ELISA Kit

Catalog No.: BEK1021

Size: 96T

Range: 156 pg/ml - 10,000 pg/ml

Sensitivity < 5 pg/ml

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

Application: For quantitative detection of Cathepsin B in human serum, plasma, body fluids, tissue lysates or cell culture supernates.

Introduction

Cathepsin B is an enzymatic protein belonging to the peptidase (or protease) families. In humans, it is coded by the CTSB gene. This protein is a lysosomal cysteine protease composed of a dimer of disulfide-linked heavy and light chains, both produced from a single protein precursor. It can degrade β-amyloid precursor protein into harmless fragments. Thus, it is conceivable cathepsin B may play a pivotal role in the natural defense against Alzheimer's disease. Overexpression of cathepsin B has been associated with esophageal adenocarcinoma and other tumors.

Principle of the Assay  

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

Kit components

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