Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen)

Salacinski, P; McLean, C; Sykes, Jane E.; Clement‐Jones, Vicky; Lowry, P. J.

doi:10.1016/0003-2697(81)90703-x

Cited by 1,170 publications

(425 citation statements)

References 19 publications

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“…Unhydrolysed phospholipid was removed by extraction into chloroform and the amount of inositol phosphate which remained in the upper phase was determined by measurement of organic phosphorus or radioactivity (8), Fifty micrograms of purified PI-PLC (1.5 nmol) were iodinated with I mCi of ['--''I]-iodidc (Amersham) using iodogcn technique according to Salacinski etal. (9). Free iodide was removed from iodinated PI-PLC on a PDIO (Pharmacia) Sephadex G25 column run in phosphate bufiered saline (PBS).…”

Section: Methodsmentioning

confidence: 99%

Antibody against Bacillus thuringiensis phosphatidylinositol‐phospholipase C: Some examples of its potential uses

Théveniau¹,

Malapert²,

Rougon³

1990

Immunology & Cell Biology

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SummaryWe have purified the phosphatidylinositol-phospholipase C enzyme from the bacterium Bacillm thuringiensis. This enzyme is able to release in soluble form molecules which are anchored lo membranes via a glycan-phosphatidylinositol group. It exhibits a molecular weight of-Vl-SS kDa. We raisedpolyclonalantiseraagainst the molecule and used them in immunoblot as well asradioimmunoassays for cnzymcdetcction. This last technique should facilitate monitoringof chromatographic steps during enzyme purification. We coupled antibodies to Sepharose beads in order to remove Ihe enzyme from incubation media. This reagent also proved to be particularly useful in control experiments designed to ascertain that the observed release of molecules is due lo the action of the phosphatidylinositol-phospholipase C enzyme and not to spontaneous release or to cleavage by nonspecific hydrolases. A search for cross-reactive molecules in other bacterial strains or mammalian tissues gave negative results. This leads to the conclusion that a great diversity exists between phosphatidylinositol-phospholipascs C, even among different bacterial strains.

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Section: Methodsmentioning

confidence: 99%

Antibody against Bacillus thuringiensis phosphatidylinositol‐phospholipase C: Some examples of its potential uses

Théveniau¹,

Malapert²,

Rougon³

1990

Immunology & Cell Biology

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“…Two weeks later, ascites was recovered, and antibodies were purified by chromatography on DEAE Trisacryl M (18). Purified antibodies were labeled with ' "1 using the IODOGEN method (19).…”

Section: Methodsmentioning

confidence: 99%

Osteoarthritis‐related changes in human articular cartilage

Vilamitjana-Amedee

Harmand

1990

Arthritis & Rheumatism

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We report the use of immunologic methods for detecting specific alterations in human osteoarthritic cartilage. Monoclonal antibodies with specificities for proteoglycans and link proteins have been used to define immunogenic sites in these molecules. We have identified 1 site in the protein core, 1 site in the link proteins, and another site that is common to both proteoglycan and link proteins, which are not modified in osteoarthritic cartilage. In addition, an antigenic determinant in link proteins that is altered in osteoarthritic cartilage has been identified. These results suggest that the structure of the ternary complex (hyaluronic acid binding region-link proteins-hyaluronic acid) could be altered in osteoarthritic cartilage. These modifications may be due to a genetic defect or to partial enzymatic degradation of these sites.In previous studies, we have shown differences between the noncollagenous proteins extracted from normal and osteoarthritic articular cartilage (1). These proteins, particularly proteoglycans and structural glycoproteins, were extracted from the osteoarthritic cartilage using lower ionic strength conditions than were used for normal cartilage. Two proteoglycan monomer populations with different molecular weights were identified in osteoarthritic cartilage. They also differed in molecular weight from the 3 populations identified in cartilage from normal age-matched do-

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“…Assay sensitivity (90% B/B 0 ) was 0.5 6 0.12 ng per tube. Purified ovine GH (NIDDK-oGH) was used for radioiodination according to Salacinsky et al (1981); specific activity was 27.6 mCi/mg. The same preparation (NIDDKoGH) was used as the reference standard.…”

Section: Shearing Stress and Hormonal Responsementioning

confidence: 99%

The effect of shearing procedures on blood levels of growth hormone, cortisol and other stress haematochemical parameters in Sarda sheep

Carcangiu

Vacca

Parmeggiani

et al. 2008

Animal

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The aim of this research was to investigate how growth hormone (GH) cortisol and some haematochemical parameters could be modified by the stress caused by the stages of shearing in Sarda breed sheep. Five groups of 10 sheep each were formed. Group A, only separated from the flock; Group B, only tied; Group C, both tied and shorn (animals in these three groups were ewe lambs shorn for the first time); Group D, adult females both tied and shorn; and Group E, adult entire males both tied and shorn (animals in these two groups had been shorn previously). Five blood samples were taken from each animal: the day before treatment (first sample); at the start of the treatment (second sample); in the middle of shearing for Groups C, D and E, 10 min after separation in Group A and 10 min after tying in Group B (third sample); at the end of treatment (fourth sample); and on the day after treatment (fifth sample). Plasma GH levels showed a decrease (P , 0.01) in Groups A, B, C and D during treatment (third and fourth samples), while Group E only at the end of shearing (fourth sample). In the third sample, the highest GH levels were recorded for Group E (P , 0.05), while it was recorded in the fourth sample for Groups A and E (P , 0.05). Cortisol levels showed a clear increase (P , 0.01) in all groups during treatment, but Group A showed a decrease in the fourth sample in comparison to the third sample. Males in the second, third and fourth sample and Group A only in the fourth sample showed lower cortisol levels when compared with the other groups (P , 0.05). Plasma glucose levels showed an increase (P , 0.01) in all groups during treatment but Groups B, C and E showed the highest values (P , 0.05). Magnesium (Mg) showed an increase in all groups in the third and fourth sample, while sodium (Na), in the same samples, only in Groups B, C and D. Potassium (K) values showed a significant decrease (P , 0.05) only in Groups C and D at the end of shearing. These results show that GH secretion is influenced by all the stress procedure: separation, tying and shearing. Shearing, even if necessary for animals, causes a significant change of the blood parameters involved in the stress response.

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Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen)

Cited by 1,170 publications

References 19 publications

Antibody against Bacillus thuringiensis phosphatidylinositol‐phospholipase C: Some examples of its potential uses

Antibody against Bacillus thuringiensis phosphatidylinositol‐phospholipase C: Some examples of its potential uses

Osteoarthritis‐related changes in human articular cartilage

The effect of shearing procedures on blood levels of growth hormone, cortisol and other stress haematochemical parameters in Sarda sheep

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