Human Kallikrein 3 ELISA Kit

Catalog No.: BEK1129

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 Kallikrein 3 in human serum, body fluids, tissue lysates or cell culture supernatants.

Introduction

Prostate-specific antigen (PSA), also known as gamma-seminoprotein or kallikrein-3 (KLK3), is a single-chain glycoprotein with a molecular mass of about 33 kD that may function normally in the liquefaction of seminal coagulum. The PSA gene spans about 6 kb and contains 5 exons. It is a member of the kallikrein-related peptidase family and is secreted by the epithelial cells of the prostate gland. PSA is produced for the ejaculate, where it liquefies semen in the seminal coagulum and allows sperm to swim freely. It is also believed to be instrumental in dissolving cervical mucus, allowing the entry of sperm into the uterus.

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-Kallikrein 3 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-Kallikrein 3 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 Kallikrein 3 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of Kallikrein 3 can be calculated.

Kit components

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