Specificity of Antibodies Synthetized by Immunocytes as Detected by Immunoenzyme Techniques

Avraméas, S; Taudou, B; Ternynck, Thérèse

doi:10.1159/000230403

Cited by 36 publications

(5 citation statements)

References 8 publications

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“…One filter from each pair was washed exhaustively in 50 mM Tris-HCI (pH 7.5) and in 0.5 M NaCl. All filters were stained for PhoA activity [12] with distilled water and resuspended in 50 mM Tris-HCI (pH 7.5), prior to use. After incubation of the protein samples with Avicel at 0°C for 1 h, the Avicel was collected by centrifugation.…”

Section: 3mentioning

confidence: 99%

Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose

et al. 1989

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Endoglucanase CenA of Cellulomonas fimi comprises an N‐terminal cellulose‐binding domain and a C‐terminal catalytic domain joined together by a sequence of 23 proline and threonine residues (the Pro‐Thr box). The domains function independently when separated by proteolysis. TnphoA has been used to generate cenA′‐′phoA fusions. CenA′‐′PhoA fusion polypeptides which contain the entire cellulose‐binding domain of CenA bind to cellulose, allowing their purification from periplasmic extracts in a single, facile step. This result has implications for purification or immobilisation of chimeric proteins on a cheap cellulose matrix.

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Section: 3mentioning

confidence: 99%

Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose

et al. 1989

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“…(2) Alkaline-phosphatase-labeled antibodies against immunoglobulins (500 ,g/ml) for 2 hr. Slides were then washed and incubated with the specific substrate of alkaline phosphatase (naphthol-AS MX and Fast Red) for 30 min (18) T Determined by subtraction of percent Id +Ab-from percent Id+.…”

Section: Methodsmentioning

confidence: 99%

Similar Idiotypes in Antibody-Forming Cells and in Cells Synthesizing Immunoglobulins Without Detectable Antibody Function

Cazenave

Ternynck

Avraméas

1974

Proc. Natl. Acad. Sci. U.S.A.

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The occurrence of immunoglobulins with and without antibody specificity and with and without idietypic specificity was studied, by use of enzyme-labeled antigen and antibodies, in lymph node cells of rabbits immunized with horse-radish peroxidase and hen ovalbumin. Some cells, containing immunoglobulins without detectable antibody function, were shown to contain idiotypes similar to those found in antibody-producing cells.Idiotypy of antibodies has been defined as their property of possessing different antigenic specificities, both among antibodies of one individual against distinct antigens and among antibodies of different individuals (or perhaps of different groups of individuals) against the same antigen (1, 2). The first observations of this property were made almost simultaneously in humans (3) and in rabbits (4). Further observations made in rabbits have shown that similar idiotypic specificities may be common to antibodies directed against a protein and to immunoglobulins apparently devoid of antibody function against protein (5; 6).In the present work, cells containing immunoglobulins with and without antibody specificity and with and without idiotypic specificities were studied by use of enzyme-labeled antigens or antibodies. The present experiments apparently confirm the results already obtained with circulating immunoglobulins, and further emphasize the close relationship existing between nonspecific immunoglobulin and antibody after specific antigenic stimulation (7)(8)(9)(10)(11)

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“…After IEF proteins were transferred to nitrocellulose sheets by electroblotting for 5 h at 35-40 V in a solution consisting of 0.7% acetic acid and 20% methanol (Towbin et al, 1979). Immune fixation was carried out by a modification (Pintar et al, 1983) of the method of Avrameas et al (1971). Incubations of nitrocellulose were at 37"C, first with a 1:1,000 dilution of antiserum to COMT or a 1:200 dilution of nonimmune serum, then with a 1:400 dilution of goat anti-rabbit immunoglobulin conjugated to alkaline phosphatase (Polysciences, Inc.).…”

Section: Gel Electrophoresis and Immune Fixationmentioning

confidence: 99%

Soluble and Particulate Forms of Rat Catechol‐O‐Methyltransferase Distinguished by Gel Electrophoresis and Immune Fixation

Grossman

Creveling

Rybczynski

et al. 1985

Journal of Neurochemistry

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Catechol-O-methyltransferase (COMT) was visualized in homogenates and subcellular fractions of rat tissues, including liver and brain, by gel electrophoresis, electrophoretic transfer of proteins to nitrocellulose (Western blotting), and immune fixation with antiserum to highly purified soluble rat liver COMT. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of all tissue homogenates examined revealed three major immune-specific proteins with apparent molecular weights 23,000, 26,000, and 66,000 (23K, 26K and 66K). Centrifugation of homogenates at 100,000 X g for 60 min resulted in the enrichment of the 26K species protein in the pellet whereas the 23K and 66K proteins were the predominant forms in the supernatant. The 66K protein appeared in variable amounts depending on the tissue being examined and the length of transfer of protein and is assumed to be an "aggregate" of the smaller form(s). The 26K protein was essentially the only immunoreactive species seen in a purified preparation of rat liver outer mitochondrial membrane. Isoelectric focusing (IEF) under denaturing conditions and two-dimensional gel electrophoresis of brain and liver fractions showed that the 23K protein was resolved into three bands of pI 5.1, 5.2, and 5.3, whereas the 26K protein had a pI of 6.2. Analysis of COMT activity in slices from nondenaturing IEF gels indicated that the pI 5.1-5.3 species are biologically active; the pI 6.2 species could not be detected under these conditions. COMT activity was demonstrated, however, in outer mitochondrial membranes from rat liver, which contain predominantly the 26K, pI 6.2 immunoreactive species. The major form of COMT in all rat tissues examined is "soluble" with an apparent Mr of 23K and a pI of 5.2. The nature of the modifications giving rise to pI 5.1 and 5.3 forms of this enzyme are not clear, nor is the relationship between the 23K and 26K forms. Further studies are needed to elucidate the relationship of immunoreactive forms of COMT to each other, their intracellular location, and their functional significance.

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Specificity of Antibodies Synthetized by Immunocytes as Detected by Immunoenzyme Techniques

Cited by 36 publications

References 8 publications

Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose

Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose

Similar Idiotypes in Antibody-Forming Cells and in Cells Synthesizing Immunoglobulins Without Detectable Antibody Function

Soluble and Particulate Forms of Rat Catechol‐O‐Methyltransferase Distinguished by Gel Electrophoresis and Immune Fixation

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