Human Progastrin-releasing peptide, ProGRP ELISA Kit

Catalog No.: BEK1255

Size: 96T

Range: 31.2 pg/ml - 2000 pg/ml

Sensitivity 2 pg/ml

Storage and Expiration: Store at 4℃ for 6 months, or at -20℃ for one year. Store HRP conjugated antibody at 4℃ for one year.

Application: For quantitative detection of human ProGRP in serum, plasma, cell culture supernatants or tissue homogenate.

Introduction

Progastrin-releasing peptide (proGRP) is a recently identified biomarker of small cell lung cancer (SCLC). Plasma proGRP level could be a useful diagnostic and therapeutic monitoring biomarker for patients with SCLC and the initial level may help with SCLC tumor staging. ProGRP was highly expressed in NCI-H446 cells and xenograft tissue. It could be used as a marker for quality control in sample processing and storage in biobanks. Korse CM1 et al study concluded that a high-risk proGRP level (more than twice the upper reference value) in patients with well-differentiated neuroendocrine tumours, WDNETs is a strong indication for a primary tumour in the lung. Besides CgA, proGRP is a complementary tumour marker for prognosis and treatment monitoring in patients with neuroendocrine tumour. 

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-ProGRP monoclonal antibody was pre-coated onto 96-well plates. The standards and test samples were added to the wells and any ProGRP present is bound by the immobilized antibody. After washing away any unbound substances, the HRP conjugated anti-ProGRP monoclonal antibody was added to wells as detection antibodies. Following a wash to remove any unbound antibody-enzyme reagent, the TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding stop solution. The density of yellow is proportional to the ProGRP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of ProGRP can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-human ProGRP antibody
2. Lyophilized human ProGRP standards: 2 tubes (10 ng / tube)                  
3. Sample / Standard diluent buffer: 30 ml
4. HRP conjugated anti-human ProGRP antibody (Concentrated): 130 μl. Dilution: 1:100 (Store at 4℃, do NOT store it at -20℃)
5. Antibody diluent buffer: 12 ml
6. TMB substrate: 10 ml
7. Stop solution: 10 ml
8. Wash buffer: 30 ml (25x). Dilution: 1:25

Note: Reconstitute standards and test samples with Kit Component 3.

Material Required But Not Provided

1. Microplate reader (wavelength: 450nm)
2. Precise pipette and disposable pipette tips
3. Deionized or distilled water
4. Automated plate washer
5. Beakers, flasks or cylinders, etc.
6. Statistical calculator
7. Plate cover
8. Absorbent filter papers or absorbent material
9. ELISA shaker