1995
DOI: 10.1007/bf00298435
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Immunoblotting studies on artifactual contamination of enamel homogenates by albumin and other proteins

Abstract: The reason for the presence of albumin and other serum, cytoskeletal, cytosolic, and extracellular matrix proteins in enamel fractions was investigated by immunoblotting using homogenates prepared from freeze-dried and freshly dissected rat incisors, and antibodies capable of resolving at least 1 ng of the primary antigen. The data indicated that most of the 16 antibodies examined in this study reacted with antigens present only within "cell" homogenates (enamel organ cells + adhering labial connective tissue … Show more

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Cited by 30 publications
(31 citation statements)
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“…Other than amelogenin, enamelin and ameloblastin, a number of other proteins have been identified in the developing and mature enamel. These findings have initiated debates on the significance of these other proteins to the development and final form of the mature enamel organ [Kinoshita, 1979;Limeback et al, 1989;Strawich and Glimcher, 1990;Chen et al, 1995]. Enamel mineralization and maturation occur rapidly in a biological time frame, and it has been discussed previously that biological molecules (such as serum proteins, lipids and proteins derived from Tomes' processes and more general extracellular proteins) cannot be readily eliminated from the enamel matrix environment by existing proteases [Zeichner-David et al, 1997;Paine et al, 2000a].…”
Section: Discussionmentioning
confidence: 99%
“…Other than amelogenin, enamelin and ameloblastin, a number of other proteins have been identified in the developing and mature enamel. These findings have initiated debates on the significance of these other proteins to the development and final form of the mature enamel organ [Kinoshita, 1979;Limeback et al, 1989;Strawich and Glimcher, 1990;Chen et al, 1995]. Enamel mineralization and maturation occur rapidly in a biological time frame, and it has been discussed previously that biological molecules (such as serum proteins, lipids and proteins derived from Tomes' processes and more general extracellular proteins) cannot be readily eliminated from the enamel matrix environment by existing proteases [Zeichner-David et al, 1997;Paine et al, 2000a].…”
Section: Discussionmentioning
confidence: 99%
“…Immunoglobulins were extracted from the yolks by polyethylene glycol precipitation. The specificity of the antibodies was verified by immunoblotting against the respective immunogens, rat serum albumin (Sigma) and HC1 bone extract (see above) as described in Chen et al (1995). Briefly, the proteins were separated by electrophoresis on 12% SDS-polyacrylamide slab gels and transferred onto 0.45 p,m nitrocellulose membranes (Xymotech Biosystems, Montreal, Quebec, Canada) at 4°C and 4,000 mA constant current for 2 hours in a Genus Electro-transfer unit (IDEA Scientific Company, Minneapolis, MN) as described by Towbin et al (1979).…”
Section: Preparation Of Chicken Egg Yolk Anti-ratmentioning
confidence: 99%
“…51 Proteins were subsequently transferred onto 0.45 mm nitrocellulose membranes (Mandel, Ontario, Canada) and probed with anti-AMBN antibody (1:1000; courtesy of Dr PH Krebsbach 52 ), anti-BSP antibody (1:500; LF-100; courtesy of Dr LW Fisher, NIDCR, NIH, Bethesda, MD, USA) and anti-OPN antibody (1:500; LF-123; courtesy of Dr LW Fisher) as described previously. 53 Antibody binding was revealed with a secondary goat anti-rabbit antibody conjugated to alkaline phosphatase (1:1000; Sigma, Oakville, Ontario, Canada) using p-nitroblue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl phosphate as substrates (Amersham Biosciences, Quebec, Canada).…”
Section: Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis Anmentioning
confidence: 99%