Human Fatty acid-binding protein, heart, HFABP / FABP3 ELISA Kit

Catalog No.: BEK1254

Size: 96T

Range: 15.6 pg/ml - 1000 pg/ml

Sensitivity 1 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 HFABP in serum, cell culture supernatants or tissue homogenate.

Introduction

Heart-type fatty acid binding protein (hFABP) also known as mammary-derived growth inhibitor is a protein that in humans is encoded by the FABP3 gene. The serum HFABP level is an effective biomarker for detecting AKI in non-surgical intensive care patients. HFABP expression is significantly higher in in-stent restenosis, ISR tissue than in non-ISR tissue from diabetic and nondiabetic minipigs. Overexpression of HFABP induces multiple pathway-related promotion of inflammation, growth and migration in vascular SMCs, suggesting a potential role in coronary artery ISR. hFABP is a rapid marker of perioperative myocardial damage and peaks earlier than CKMB or TnI. The kinetics of marker proteins in serial samples immediately after reperfusion is more suitable for the detection of perioperative myocardial infarction than a fixed cut-off level. 

Principle of the Assay  

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-HFABP monoclonal antibody was pre-coated onto 96-well plates. The standards and test samples were added to the wells and any HFABP present is bound by the immobilized antibody. After washing

away any unbound substances, the HRP conjugated anti-HFABP 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 HFABP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of HFABP can be calculated.

Kit components

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