Two‐Dimensional electrophoresis of human serum proteins following acute myocardial infarction

Abstract

Serum proteins associated with acute myocardial infarction (AMI) have been monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and high resolution two-dimensional electrophoresis (2-DE) under nonreducing conditions. Proteins a, b, c (Mr 13,000; pI6.2, 6.7 and 7.5, respectively) and e(Mr27,000; pI5.2) appear simultaneously approximately 30 h after infarction, reach maximum intensity after 48 h and progressively decline thereafter. Protein d (Mr15,000; pI7-8.5; identified as hemoglobin) sometimes appears within 18 h of infarction. Proteins a-c are not detected in the 2-DE patterns of healthy myocardium, infarcted myocardium, pectoral muscle or tongue, but e is present in all and tentatively identified as myosin light chain. Other myocardial proteins which are either reduced in amount following infarction or more specifically associated with myocardium than pectoral muscle are not detected in the serum of AMI patients. Analysis of unconcentrated urine by SDS-PAGE and silver staining does not reveal proteins specific to AMI.

The simplified technique of high resolution two‐dimensional polyacrylamide gel electrophoresis: Biomedical applications in health and disease

“…4). The other abnormal polypeptides (M, 13 000; pZ6.2,7.2, and 7.5) are probably isoform; of apo-serum amyloid A protein, an acute-phase reactant [19,20,281.…”

“…9A) and are thought to provide a "first line of defense" against dietary toxins by complexing polyphenolics [71]. They show a variable distribution in different individuals [72,73] and appear as diffuse zones, particularly following high resolution 2-D PAGE (Fig. 9A).…”

Electrophoretic analysis of stimulated cat parotid saliva

“…Duplicate sample aliquots were prepared without 2-mercaptoetha-no1 ("non-reducing" conditions) and also with a range of SDS concentrations (0.5%-5.0% w/v). The sample mixtures were heated to 95°C for 10 min, loaded in agarose wells precast on the upper surface of 6-24% w/v polyacrylamide gradient gels (75 X 75 X 3 nun) and electrophoresed, without a stacking gel, at 70 mA/gel for lh in precooled (4°C) 0.025 M Tris, 0.2 M glycine containing 0.1% w/v SDS [2,3].in aqueous 20% w/v trichloroacetic acid and the proteins stained at 60°C for 30 min in aqueous 50% v/v methanol/lO% acetic acid containing 0.1% w/v Serva Blue R prior to destaining in aqueous 5% v/v methanol /7% acetic acid and densitometry (Ultrascan XL Laser Densitometer, LKB, Milton Keynes, UK).The major cat parotid salivary protein appeared as a monomer (Mr 30,000) upon SDS-PAGE under "reducing" conditions but as a dimer (Mr 60,060) under "non-reducing" conditions. High resolution two-dimensional electrophoresis under " n o n -r e d u c i n g " c o n d i t i o n s revealed only the monomersuggesting a non-covalent association.…”

“…Duplicate sample aliquots were prepared without 2-mercaptoetha-no1 ("non-reducing" conditions) and also with a range of SDS concentrations (0.5%-5.0% w/v). The sample mixtures were heated to 95°C for 10 min, loaded in agarose wells precast on the upper surface of 6-24% w/v polyacrylamide gradient gels (75 X 75 X 3 nun) and electrophoresed, without a stacking gel, at 70 mA/gel for lh in precooled (4°C) 0.025 M Tris, 0.2 M glycine containing 0.1% w/v SDS [2,3].…”