Description

Nori Rat H-FABP ELISA Kit Summary

Alternative names for H-FABP: Heart-type fatty acid binding protein, FABP3, FABP11, MDG1

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Heart-type fatty acid binding protein (H-FABP) also known as mammary-derived growth inhibitor, is a protein that in humans is encoded by the FABP3 gene.^[1] Its function is to arrest growth of mammary epithelial cells. It is also a candidate tumor suppressor for human breast cancer. H-FABP is a small cytoplasmic protein (15 kDa) released from cardiac myocytes following an ischemic episode.^[2] Like the nine other distinct FABPs, H-FABP is involved in active fatty acid metabolism where it transports fatty acids from the cell membrane to mitochondria for oxidation.^[2]  H-FABP is a sensitive biomarker for myocardial infarction (MI)^[3] and can be detected in the blood within 1-3 hours of the pain. H-FABP is 20 times more specific to cardiac muscle than myoglobin,^[4]  H-FABP is recommended to be measured with troponin to identify MI and acute coronary syndrome in patients with chest pain. H-FABP measured with troponin shows increased accuracy and sensitivity of 20.6% over troponin at 3-6 hours following chest pain onset.^[5]  It rapidly release into plasma after myocardial injury – 60 minutes after an ischemic episode,^[6] and has tissue specificity. Measuring H-FABP in combination with troponin has a negative predictive value of 98%, could be used to identify those not suffering from MI at the early time point of 3-6 hours post chest pain onset.^{[5] [6]} Alongside D-dimer, NT-proBNP and peak troponin T, H-FABP was the only cardiac biomarker that proved to be a statistically significant predictor of death or MI at one year. Patients who were TnI negative but H-FABP positive had 17% increased risk of all cause mortality within one year compared to those patients who were TnI positive but H-FABP negative.^[7]  H-FABP has been proven to significantly predict 30 day mortality in acute pulmonary embolism.^[8] H-FABP is more effective than Troponin T in risk stratifying Chronic Heart Failure patients.^[9]

References

1. Phelan CM, et al. (1996). Genomics34(1): 63–8.
2. Kleine AH, et al. (1992). Molecular and Cellular Biochemistry116(1-2): 155–62.
3. Tanaka T, et al. (1991). Clinical Biochemistry24 (2):
4. Ghani F, et al. (2000). Clinical Chemistry46 (5): 718–9.
5. Glatz JF, et al. (1994).  British Heart Journal71 (2): 135–40.

1. Van Nieuwenhoven FA, et al. (1995). Circulation 92 (10): 2848–54.
2. Viswanathan K, et al. (2010). Journal of the American College of Cardiology 55: 2590–8.
3.  Kaczyñska A, et al. (2006). Clinica Chimica Acta; Intl J of Clinical Chem 371 (1-2): 117–23.
4. Niizeki T, et al. (2007). Journal of Cardiac Failure 13 (2): 120–7.

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