Cat. No.
MAK-0075
Product Name
Dog H-FABP
Storage
4°C
Species
Dog
Instructions
All reagents should be allowed to reach room temperature before use.
Introduction
Fatty acid-binding proteins are cytoplasmic proteins of approximately 15 kDa involved in fatty acid transport and metabolism. The isoform expressed in heart (H-FABP) is highly abundant, representing 10-20 mol % of total cytoplasmic protein. HFABP is released into blood after cardiac injury and provides a sensitive biomarker of myocardial necrosis in several species.1-5 HFABP is expressed in both cardiac and skeletal muscle. When used as a cardiac biomarker, it is therefore important to rule out skeletal muscle injury. In the absence of cardiac injury, H-FABP can also be used as a biomarker of skeletal muscle damage. As shown in the figure below, we found that H-FABP levels peaked two hours after cardiac ischemia-reperfusion in dogs.
Principle Of The Assay
The ELISA uses two H-FABP antibodies. One is used for solid phase immobilization (microtiter wells). The other is conjugated to horse radish peroxidase (HRP) and used for detection. Standards and diluted samples are incubated in the microtiter wells with HRP conjugate for one hour. This results in H-FABP molecules being sandwiched between the immobilization and detection antibodies. The wells are then washed to remove unbound HRP conjugate. TMB is added and incubated for 20 minutes. If H-FABP is present a blue color develops. Color development is stopped by addition of Stop solution, changing the color to yellow, and absorbance is measured at 450 nm. The concentration of H-FABP is proportional to absorbance and is derived from a standard curve.
