Human Procalcitonin, PCT ELISA Kit

Catalog No.: BEK1246

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 PCT in human serum, plasma, tissue homogenates or cell culture supernatants.

Introduction

Procalcitonin (PCT) is a peptide precursor of the hormone calcitonin, the latter being involved with calcium homeostasis. It is composed of 116 amino acids and is produced by parafollicular cells (C cells) of the thyroid and by the neuroendocrine cells of the lung and the intestine. Measurement of procalcitonin can be used as a marker of severe sepsis caused by bacteria and generally grades well with the degree of sepsis, although levels of procalcitonin in the blood are very low. PCT has the greatest sensitivity (85%) and specificity (91%) for differentiating patients with systemic inflammatory response syndrome (SIRS) from those with sepsis, when compared with IL-2, IL-6, IL-8, CRP and TNF-alpha. It is a biomarker of bacterial infection which may help gauge the severity and prognosis of patients with CAP. In addition to clinical predictors, PCT may assist in decisions pertaining to timing of discharge from hospital and the discontinuation of antibiotics.

Principle of the Assay  

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

Kit components

1. One 96-well plate pre-coated with anti-Human PCT antibody
2. Lyophilized Human PCT standards: 2 tubes (10 ng/tube)                     
3. Sample / Standard diluent buffer: 30 ml
4. Biotin conjugated anti-Human PCT antibody (Concentrated): 130 μl. Dilution: 1:100
5. Antibody diluent buffer: 12 ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130 μl. Dilution: 1:100
7. ABC diluent buffer: 12 ml
8. TMB substrate: 10 ml
9. Stop solution: 10 ml
10. Wash buffer (25X): 30 ml

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