Human Pro-Cathepsin B ELISA Kit

Catalog No.: BEK1317

Size: 96T

Range: 156 pg/ml - 10,000 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 Pro-Cathepsin B in human serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Introduction

Cathepsin B belongs to a family of lysosomal cysteine proteases known as the cysteine cathepsins and plays an important role in intracellular proteolysis. In humans, cathepsin B is encoded by the CTSB gene. Cathepsin B is upregulated in certain cancers, in pre-malignant lesions, and in various other pathological conditions. The CTSB gene is located at chromosome 8p22, consisting of 13 exons. The promoter of CTSB gene contains a GC-rich region including many SP1 sites, which is similar to housekeeping genes. At least five transcript variants encoding the same protein have been found for this gene.

Principle of the Assay  

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

Kit components

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