Méthode immuno-électrophorétique d'analyse de mélanges de substances antigéniques

Grabar, P; Williams, Curtis A.

doi:10.1016/0006-3002(55)90320-6

Cited by 649 publications

(129 citation statements)

References 6 publications

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“…The immunoelectrophoretic plates were a modification of the technique of Grabar & Williams (1955). A sample (10 g.) of Noble special agar (Difco Laboratories, Detroit, Michigan, U.S.A.) was washed and made up to 500ml.…”

Section: Methodsmentioning

confidence: 99%

Immunological Studies of Amination Deficient Strains of Neurospora crassa

Roberts

Pateman

1964

Journal of General Microbiology

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SUMMARYA survey has been made of the immunological relationships between glutamate dehydrogenase (GDH) and the related proteins of the amination deficient (am) mutants of Neurospora crassa. Serum against GDH was prepared in rabbits which had been injected with crude extracts of wild type N. crwsa. The effect of this serum on the activity of normal GDH and the activity of activated mutant proteins was studied. The techniques of double diffusion in agar gel and of immunoelectrophoresis were used to study the relationships between the wild-type GDH and mutant forms of the protein, The identification of the relevant precipitation line on these gel plates was' confirmed by the use of a specific stain for the enzyme antibody complex. The relationship between the antigenic and enzymic sites in the wild-type protein is discussed.

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Section: Methodsmentioning

confidence: 99%

Immunological Studies of Amination Deficient Strains of Neurospora crassa

Roberts

Pateman

1964

Journal of General Microbiology

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“…The moles of carboxylic group liberated during the reaction were measured by the method of Grassman & Heyde (1929) and the percentage hydrolysis is given by the relation: moles of carboxylic group liberated x 100 moles of substrate employed Detection of cathep8in8 D and E after electrophore8i8in agar. Different enzyme preparations were subjected to electrophoresis in agar gel according to the technique of Grabar & Williams (1955) under the following conditions: volume of preparation used, 0-05 ml. ; plate of 1% agar, 10-5 cm.…”

Section: Methodsmentioning

confidence: 99%

The purification and properties of a proteolytic enzyme, rabbit cathepsin E, and further studies on rabbit cathepsin D

Lapresle¹,

Webb²

1962

Biochemical Journal

126

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“…30 Sheep anti-human apoB and apoA-I were added to the wells to identify the respective apolipoproteins. The gels were stained with Coomassie blue R-250.…”

Section: Lmmunoelectrophoresis Of Plasma and Plaque Ljpoproteinsmentioning

confidence: 99%

Triglyceride-rich lipoproteins isolated by selected-affinity anti-apolipoprotein B immunosorption from human atherosclerotic plaque.

Rapp

Lespine

Hamilton

et al. 1994

Arterioscler Thromb

310

185

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We isolated and characterized immunoreactive apolipoprotein B (apoB) -containing lipoproteins from human atherosclerotic plaque and plasma to determine whether verylow-density lipoprotein (VLDL) can enter and become incorporated into the atherosclerotic lesion and how plaque apoBcontaining lipoproteins differ from apoB-containing lipoproteins isolated from plasma. Atherosclerotic plaques were obtained during aortic surgery and processed immediately. Lipoproteins were extracted from minced plaque in a buffered saline solution (extract A). In selected cases a second extraction was done after plaque was incubated with collagenase (extract B). Lipoproteins were then isolated from the extracts by anti-apoB immunosorption and separated into VLDL+intermediate-density lipoprotein (IDL) (d< 1.019 g/mL) and low-density lipoprotein (LDL) (1.019«f< 1.070 g/mL) fractions by ultracentrifugation. The VLDL+IDL fractions from plaque contained more than one third of the total apoB-associated lipoprotein cholesterol in both extracts A and B. The lipid composition of VLDL+IDL in both extracts was related to that of plasma VLDL+IDL By electron microscopy L ipoprotein infiltration into the artery wall is an essential event in the pathogenesis of atherosclerosis, but little is known about the process of lipoprotein infiltration and eventual entrapment in plaque. Even the identity of the lipoproteins involved has not been conclusively determined. It is generally agreed that low-density lipoproteins (LDL) have access to the artery wall, but very-low-density lipoproteins (VLDL) also may infiltrate the artery wall and directly contribute to atherogenesis. VLDL, particularly VLDL remnants, certainly have atherogenic potential.1 -2 Particles resembling VLDL remnants can be taken up by macrophages to produce foam cells, 3 -3 stimulate endothelial cells to express a monocyte-specific chemotactic factor, 6 and increase monocyte adherence to the endothelium. 7 Furthermore, varying amounts of labeled © 1994 American Heart Association, Inc.mean particle diameters of VLDL+IDL from extracts A and B were 9% and 23%, respectively, greater than VLDL+IDL diameters from plasma. Mean diameters of LDL from extracts A and B were 11% and 31% greater than LDL diameters from plasma. The apoE-apoB ratio of extract A VLDL+IDL was nearly twice that of plasma VLDL+IDL and severalfold higher than that of extract A LDL. Immunoblots of both VLDL+IDL and LDL from extract A demonstrated minimal fragmentation of apoB. These results demonstrate that (1) VLDL, VLDL remnants, or both, can enter human atherosclerotic plaque and become bound to the connective tissue matrix; (2) apoB-containing lipoproteins from plaque differ from plasma lipoproteins in size and apoE content; and (3) there appears to be minimal fragmentation of buffer-extractable apoB in plaque lipoproteins. (Arterioscler Thromb.

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Méthode immuno-électrophorétique d'analyse de mélanges de substances antigéniques

Cited by 649 publications

References 6 publications

Immunological Studies of Amination Deficient Strains of Neurospora crassa

Immunological Studies of Amination Deficient Strains of Neurospora crassa

The purification and properties of a proteolytic enzyme, rabbit cathepsin E, and further studies on rabbit cathepsin D

Triglyceride-rich lipoproteins isolated by selected-affinity anti-apolipoprotein B immunosorption from human atherosclerotic plaque.

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