Improved reaction buffers for solid-phase enzyme immunoassay without interference by serum factors

Kato, Kanefusa; Umeda, Yumiko; Suzuki, Fujiko; Kosaka, Atsuko

doi:10.1016/0009-8981(80)90043-1

Cited by 43 publications

(11 citation statements)

References 4 publications

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“…Procedures of the immunoassay were almost the same as those described for the assay of aldolase-C (Haimoto and Kato, 1986). In brief, a piece of polystyrene ball with antibodies was incubated with the standard phosphorylase or samples at 30°C for 5 h with shaking in a final volume of 0.5 ml of 10 mM Tris-HC1 buffer, pH 7.0, containing 0.3 M NaCl, 1 mMMgC12, 0.1% bovine serum albumin, 0.5% protease-treated gelatin (Kato et al, 1980), and 0.1% NaN3 (buffer G). After being washed, the ball was incubated at 4°C overnight with 2 milliunits (expressed as units of galactosidase activity; 1 unit = I pmol of product/min under the assay conditions) of the galactosidase-labeled antibodies in 0.2 ml of 10 mM sodium phosphate buffer, pH 7.0, containing 0.1 M NaCI, 1 m M MgClz, 0.1 % bovine serum albumin, and 0.1 % NaN3.…”

Section: Immunoassay System and Assay Proceduresmentioning

confidence: 99%

Human Brain‐Type Glycogen Phosphorylase: Quantitative Localization in Human Tissues Determined with an Immunoassay System

Kato

Shimizu

Kurobe

et al. 1989

Journal of Neurochemistry

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Glycogen phosphorylase (EC 2.4.1.1) from human brain tissue was purified to homogeneity. Antisera were developed in rabbits with purified phosphorylase as the immunogen. Antibodies were first affinity-purified with a column of brain phosphorylase-coupled Sepharose, and then the antibody fraction was adsorbed with a column of muscle phosphorylase-coupled Sepharose to remove antibodies reactive also with muscle phosphorylase. By using the specific antibodies, a sandwich-type immunoassay system for measurement of brain phosphorylase was prepared. The assay system consisted of polystyrene balls with immobilized antibrain phosphorylase F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The assay was sensitive and specific to the brain phosphorylase. The minimum detection limit of the assay was 0.1 ng/assay tube, and the cross-reactivity of the assay with muscle phosphorylase was less than 1%. Tissue concentrations of immunoreactive brain-type phosphorylase were estimated. The phosphorylase was present in the heart at as high a level as in the brain. The immunoreactivity for brain phosphorylase was distributed widely at a significant concentration in various peripheral tissues, such as the digestive tract, bladder, aorta, liver, and testis. Immunohistochemical localization of brain phosphorylase in the CNS revealed that the enzyme is present in most astrocytes and amyloid bodies, as well as in some neurons in the cerebral cortex and Golgi cells in the cerebellar cortex.

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Section: Immunoassay System and Assay Proceduresmentioning

confidence: 99%

Human Brain‐Type Glycogen Phosphorylase: Quantitative Localization in Human Tissues Determined with an Immunoassay System

Kato

Shimizu

Kurobe

et al. 1989

Journal of Neurochemistry

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“…A polystyrene ball with antibodies was incubated with shaking at 30°C for 5 h with a 10-pl sample or standard Goa subunit and 490 p1 of 10 m M sodium phosphate buffer (pH 7.0) containing 0.3 MNaCl, 1 mMMgC12, 0.1% BSA, 0.5% protease-treated gelatin (Kato et al, 1980), and 0.1% NaN3 (buffer GJ in a test tube (10 X 75 mm). Each ball was washed twice with 1 ml of buffer A, transferred into a fresh test tube with 200 pl of buffer A containing 2 mU of galactosidaselabeled antibody, and left standing at 4°C overnight.…”

Section: Procedures For Immunoassaymentioning

confidence: 99%

Highly Sensitive Immunoassay for the α Subunit of the GTP‐Binding Protein Go and Its Regional Distribution in Bovine Brain

Asano

Semba

Ogasawara

et al. 1987

Journal of Neurochemistry

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Antisera were raised in rabbits against the alpha subunit of a GTP-binding protein, Go. Because the antisera cross-reacted weakly with the alpha subunit of inhibitory GTP-binding protein of adenylate cyclase (Gi), they were purified with a Go alpha-coupled Sepharose column. Purified antibodies reacted only with Go alpha and did not cross-react with the Gi alpha subunit or beta gamma subunits in an immunoblot assay. Using these purified antibodies, a highly sensitive enzyme immunoassay method for the quantification of bovine brain Go alpha was developed. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimal detection limit of the assay was 0.1 fmol, or 4 pg. The assay was specific for Go alpha, and it did not cross-react with Gi alpha or beta gamma. Samples from various regions of bovine brain were solubilized with 2% sodium cholate and 1 M NaCl, and the concentrations of Go alpha were determined. Go alpha was detected in all the regions, and the highest concentration was observed in the cerebral cortex. The immunohistochemical study showed that the neuropil was rich in Go alpha.

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“…Recently, Kato 0.3 mol/1 NaCl, 1 mmol/1 MgCl2, 1 g/lNaN3, 1 g/1 bovine serum albumin and 5 g/1 protease-treated gelatin (12). Buffer A was composed of 10 mmol/1 sodium phosphate buffer (pH 7.0) containing 0.1 mol/1 NaCl, 1 mol/I MgCl2, 1 g/1 bovine serum albumin and 1 g/1 NaNî.…”

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Localization of Cu/Zn and Mn superoxide dismutase in various thyroid disorders

Iwase

Nagasaka²,

Kato³

et al. 1993

Acta Endocrinologica

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The intracellular localization of Cu/Znand Mn-superoxide dismutases (SOD), which catalyze the dismutation of superoxide radicals (O2\m=-\) to O2 and H2O2, was studied in the thyroid tissue of various thyroid disorders by an immunohistochemical technique. The concentrations of both SODs in those tissues were measured also by a sandwich enzyme immunoassay technique.Copper/zinc-SOD in thyroid tissues were identified by immunocytochemical staining in most cases of papillary carcinoma and in some cases of other thyroid disorders. In normal follicular cells this enzyme is localized in the perinuclear cytoplasm, whereas in thyroid tumor or hyperplastic follicular cells it exists homogenously in cytoplasm. Manganese-SOD stained strongly in papillary carcinoma and papillary-growing cells in the thyroid tissue of adenoma and Graves' disease. The concentrations of Cu/ Znand Mn-SOD in thyroid tumor tissues and hyperplastic follicular disorders were significantly higher than those in normal thyroid tissue when they were compared as a function of protein or deoxyribonucleic acid contents. The ratio of Mn-SOD to Cu/Zn-SOD was significantly higher only in papillary carcinoma, except for other thyroid disorders, as compared with that in the normal thyroid. In conclusion. SOD seems to be related to cell proliferation and differentiation in the thyroid follicular cell because Cu/Zn-SOD changes its localization in tumor and hyperplastic follicular cells and because the Mn-SOD concentration is increased in papillary carcinoma or papillary-growing cells.

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Improved reaction buffers for solid-phase enzyme immunoassay without interference by serum factors

Cited by 43 publications

References 4 publications

Human Brain‐Type Glycogen Phosphorylase: Quantitative Localization in Human Tissues Determined with an Immunoassay System

Human Brain‐Type Glycogen Phosphorylase: Quantitative Localization in Human Tissues Determined with an Immunoassay System

Highly Sensitive Immunoassay for the α Subunit of the GTP‐Binding Protein Go and Its Regional Distribution in Bovine Brain

Localization of Cu/Zn and Mn superoxide dismutase in various thyroid disorders

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