scholarly journals STUDIES ON ALKALINE AND ACID RIBONUCLEASES IN MAMMALIAN TISSUES IMMUNOHISTOCHEMICAL LOCALIZATION AND IMMUNOCHEMICAL PROPERTIES

1967 ◽  
Vol 15 (11) ◽  
pp. 662-673 ◽  
Author(s):  
SHIGERU MORIKAWA
Keyword(s):  
Small Intestine ◽  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Rnase A ◽  
Specific Antibodies ◽  
Mammalian Tissues ◽  
Tetramethylrhodamine Isothiocyanate ◽  
Bovine Pancreas

The distribution of alkaline and acid ribonucleases (RNases) was investigated in bovine tissues using fluorescent antibody techniques. The immunochemical properties of the bovine RNases were also examined. Specific antibodies against acid and alkaline RNase obtained by immunizing rabbits with the corresponding antigens were conjugated with fluorescein isothiocyanate or tetramethylrhodamine isothiocyanate. Anti-alkaline RNase antibody reacted against alkaline RNase-A, and anti-acid RNase antibody reacted against two different acid RNases with no cross-reactivity between alkaline and acid RNases. Phosphate-buffered 10% formalin was the best general fixative among those examined, but some differences in staining results were observed depending upon the tissue and fixative employed. Two different patterns of distribution of alkaline RNase in pancreas and kidney were observed depending upon the fixative employed. The localization of RNases in bovine pancreas, liver, small intestine, kidney, spleen, lymph node, thymus, adrenal and heart were investigated and the physiologic roles of the RNases were discussed.

scholarly journals IMMUNOHISTOCHEMICAL LOCALIZATION OF CATALASE IN MAMMALIAN TISSUES

1969 ◽  
Vol 17 (1) ◽  
pp. 30-35 ◽  
Author(s):  
SHIGERU MORIKAWA ◽  
TAKAYUKI HARADA
Keyword(s):  
Fluorescent Antibody ◽  
Bovine Liver ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Hepatic Cells ◽  
Surface Active Agents ◽  
Erythrocyte Catalase ◽  
Mammalian Tissues ◽  
Splenic Cells ◽  
Proximal Tubuli

The distribution of catalase was investigated in bovine tissues using fluorescent antibody techniques. The immunochemical properties of liver catalase were also examined. Two distinct components of liver catalase in immune system were found. Both of them possessed common antigenicity with erythrocyte catalase. No cross-reactivity was observed by immunodiffusion between catalase and the other hemoproteins such as lactoperoxidase, cytochrome c and hemoglobins. Bovine liver, pancreas, kidney, spleen and peripheral blood were examined. Catalase was located mainly in the cytoplasm of hepatic cells, acinar cells of the pancreas, epithelia of proximal tubuli, splenic cells scattered in the red pulp and some leukocytes. It was not found in any nucleus. Intracorpuscular catalase could be revealed in the erythrocytes treated with surface-active agents but not in frozen sections.

scholarly journals Presence of plasma cells binding autologous antibody during an immune response.

1979 ◽  
Vol 150 (5) ◽  
pp. 1265-1270 ◽  
Author(s):  
S Jackson ◽  
J Mestecky
Keyword(s):  
Immune Response ◽  
Human Serum Albumin ◽  
Serum Albumin ◽  
Plasma Cells ◽  
Fluorescein Isothiocyanate ◽  
Cross Reactivity ◽  
Lymphoid Tissues ◽  
Specific Antibodies ◽  
Tetramethylrhodamine Isothiocyanate ◽  
Idiotypic Antibody

Spleen and other lymphoid tissues of rabbits immunized with human serum albumin (HSA) and human lactoferrin (LF) were examined for the presence of cells forming anti-idiotype antibodies. To detect these cells, IgG, F(ab')2, or Fab' of specific antibodies were isolated, fluorochrome-tagged with tetramethylrhodamine isothiocyanate, and used as an idiotypic marker to detect splenic plasma cells that are producing anti-idiotypic antibody. By this procedure, we were able to demonstrate anti-idiotypic cells in surprisingly high numbers. For example, in six rabbits immunized with HSA for periods ranging from 36 to 542 d, the percentage of Ig-positive cells that stained with autologous idiotype ranged from 0.7 to 44; furthermore, cross-reactivity was observed among seven different anti-HSA preparations and two anti-LF antisera. The isotype of anti-idiotypic cells, determined by costaining with fluorescein isothiocyanate-labeled goat Fc-specific anti-rabbit Ig, was shown to be predominantly IgG. These findings provide evidence of the presence of plasma cells producing antibody to autologous idiotype during a vigorous immune response.

Fluorescent antibody techniques have allowed for the direct identification and enumeration of individual bacteria in environmental samples without requiring prior growth in culture media (Bahlool and Schmidt 1980, Cloete and Steyn 1988, Macario et al. 1989). The technique involves the use of specific antibodies raised against surface markers of defined pure cultures that are either labelled directly with fluorescent dye molecules or via a fluorescent secondary antibody. This approach has yielded important insights into the spatial distribution of microorganisms, but it suffers from a number of disadvantages. For example, expression of the antigen may be influenced by environmental factors; false-positive and false-negative results may be obtained due to cross-reactivity or lack of reaction; non-specific binding of antibodies may result in high levels of background fluorescence; and production of specific antibodies requires a pure culture of the organism of interest (Cloete and de Bruyn Various recombinant DNA techniques have subsequently been developed that are independent of cultivation methods (Fig. 1). These techniques provide ways of detecting and quantifying specific phylogenetic groups of microbes on 16S rDNA sequences, and relevant structural genes provide ways of monitoring microbial populations of environmental and industrial systems. In addition to these tools, a number of emerging technologies such as the use of biomarker genes are being increasingly used to monitor with great precision and accuracy the behaviour of microbes in the environment.

2003 ◽  
pp. 218-219
Keyword(s):  
Recombinant Dna ◽  
Culture Media ◽  
Specific Binding ◽  
Fluorescent Antibody ◽  
False Negative ◽  
Cross Reactivity ◽  
Phylogenetic Groups ◽  
Specific Antibodies ◽  
Negative Results ◽  
Recombinant Dna Techniques

scholarly journals IMMUNOHISTOCHEMICAL LOCALIZATION OF LACTOPEROXIDASE IN BOVINE TISSUES

1973 ◽  
Vol 21 (9) ◽  
pp. 804-811 ◽  
Author(s):  
TAKAYUKI HARADA ◽  
MITSUO BABA ◽  
SHIGERU MORIKAWA
Keyword(s):  
Mammary Gland ◽  
Fluorescent Antibody ◽  
Acinar Cells ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Sublingual Gland ◽  
Alveolar Cells ◽  
Splenic Cells ◽  
Peripheral Leukocytes ◽  
Red Pulp

Distribution of lactoperoxidase in bovine tissues was investigated by using fluorescent antibody techniques, and immunochemical properties of the enzyme were also examined. As a result of immunodiffusion, two antigenic components were found, but no cross-reactivity between lactoperoxidase and hemoproteins such as catalase, cytochrome c, hemoglobin or ferritin was observed. Lactoperoxidase was found mainly in the cytoplasm of alveolar cells of the mammary gland; in those acinar cells, which were morphologically identical to serous cells, of the sublingual gland; in acinar cells of the lacrimal gland; and also in some peripheral leukocytes and in a small number of splenic cells in the red pulp. Lactoperoxidase in the alveolar and ductal spaces and in some of the ductal epithelia of those glands was observed after ethanol or acidified ethanol fixation but not after formalin fixation. It has been demonstrated by both the present and previous work (Morikawa and Harada (1969)) that lactoperoxidase and liver-catalase were distinguishable in bovine tissues when fluorescent antibody techniques were utilized.

The fluorescent antibody test for the identification of strains of Clostridium botulinum type E

1970 ◽  
Vol 16 (10) ◽  
pp. 917-921 ◽  
Author(s):  
A. C. Emeruwa ◽  
F. E. Ashton ◽  
R. Z. Hawirko
Keyword(s):  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Specific Staining ◽  
Vegetative Cells ◽  
Carbonate Buffer ◽  
Clostridium Botulinum Type E ◽  
Homologous Strain ◽  
The Cross

A fluorescent antibody test for the identification of toxigenic and nontoxigenic type E strains of C. botulinum is described. Hyperimmune rabbit sera were prepared to cell suspensions of the D8, 070, and Beluga strains and to spores of the D8 and PM-15 strains. Fluorescein isothiocyanate immunoglobulin conjugates were prepared and absorbed with C. bifermentans and diluted to eliminate non-specific staining and cross-reactivity with related serotypes. Enzyme pretreatment of spores and washing the stained spores in carbonate buffer, pH 9.0, enhanced the sensitivity of the test. Conjugates of cell antiserum, in most cases, stained the spore bodies and not the vegetative cells of the cross-reacting strains. Conjugates of spore antiserum stained vegetative cells as well as spores but the fluorescence of the cells was eliminated by absorption of the conjugates with young cells of the homologous strain. The absorbed conjugates stained spores of 19/20 isolates from diverse geographical areas without any reduction in the intensity of fluorescence, indicating that spore antigens carry the determinant groups of type E specificity and that the common antigen is a component of the spore body. In addition, some of the cross-reacting strains showed the same degree of fluorescence as the homologous strain, suggesting that more than one antigen may be shared by these strains.

Immunohistochemical Localization of Prolactin in the Pituitary Gland of the Pigeon, Columba livia

1972 ◽  
Vol 50 (11) ◽  
pp. 1021-1025 ◽  
Author(s):  
B. A. McKeown
Keyword(s):  
Pituitary Gland ◽  
Fluorescein Isothiocyanate ◽  
Columba Livia ◽  
Immunohistochemical Localization ◽  
Frozen Sections ◽  
Specific Antibodies ◽  
Cephalic Lobe ◽  
Fresh Frozen ◽  
Pituitary Glands ◽  
Ovine Prolactin

Specific antibodies to ovine prolactin were produced in rabbits. The antisera were conjugated to fluorescein isothiocyanate and "purified" by column chromatography. Antibodies, thus labelled, were applied to fresh-frozen sections of pigeon pituitary glands. Fluorescence was observed in specific cells of the cephalic lobe. After staining, these cells were identified as the erythrosinophilic eta cells.

Evidence for presence of a reduced form of digoxin-like immunoreactive factor (dihydro-DLIF) in mammalian tissues

1996 ◽  
Vol 42 (7) ◽  
pp. 1092-1099 ◽  
Author(s):  
H M Qazzaz ◽  
S A Jortani ◽  
J M Poole ◽  
R Valdes
Keyword(s):  
Lactone Ring ◽  
Fold Increase ◽  
Cross Reactivity ◽  
Reduced Form ◽  
Molar Ratio ◽  
Endogenous Ligand ◽  
Metabolic Route ◽  
Mammalian Tissues ◽  
Spectral Absorbance ◽  
Cytochrome P 450

Abstract Digoxin-like immunoreactive factor (DLIF) from adrenal glands is an endogenous ligand structurally related to the plant-derived cardiac glycoside digoxin. Cardiac glycosides regulate the activity of the sodium pump and thus play key roles in disease processes involving regulation of ion transport. We now report the discovery of an endogenous dihydro-DLIF analogous to dihydrodigoxin. We used HPLC, ultraviolet spectrophotometry, and cross-reactivity with two antibodies, one specific for digoxin and one for dihydrodigoxin, to support the hypothesis that dihydro-DLIF contains a chemically reduced lactone ring. The spectral absorbance maximum for dihydro-DLIF is at 196 nm, identical to dihydrodigoxin. DLIF and dihydro-DLIF are 975- and 2588-fold less immunoreactive than digoxin and dihydrodigoxin for their respective antibodies. The molar ratio of dihydro-DLIF to DLIF is approximately 5.3 in bovine adrenocortical tissue and approximately 0.38 in human serum. Dihydrodigoxin (reduced lactone ring) added to microsomes isolated from bovine adrenal cortex produced a 4.5-fold increase in digoxin-like immunoreactivity (oxidized lactone ring) after 3 h of incubation. The biotransformation is likely mediated by a cytochrome P-450 NADPH-dependent process. Our findings demonstrate the presence of a dihydro-DLIF in mammals and suggest a metabolic route for synthesis of endogenous DLIF in mammalian tissue.

RADIOIMMUNOASSAY FOR 2-METHOXYOESTRONE IN HUMAN PLASMA

1979 ◽  
Vol 91 (1) ◽  
pp. 158-166 ◽  
Author(s):  
Günter Emons ◽  
Peter Ball ◽  
Gertrud v. Postel ◽  
Rudolf Knuppen
Keyword(s):  
Human Plasma ◽  
Plasma Concentrations ◽  
Cross Reactivity ◽  
Elderly Men ◽  
Specific Antibodies ◽  
Assay Procedure ◽  
Menopausal Women ◽  
Bovine Serum Albumin Conjugate ◽  
Post Menopausal

ABSTRACT A bovine serum albumin conjugate of 2-methoxyoestrone was used for the preparation of highly specific antibodies in rabbits. Cross-reactivity for catecholoestrogens and monophenolic steroids was below 0.3 %. Only 2-methoxyoestradiol cross-reacted with 44 %. An assay procedure for the determination of unconjugated and conjugated 2-methoxyoestrone in human plasma is described. The following mean plasma concentrations (pg/ml) were found (unconjugated/conjugated): children 61/1130, young men 74/1320, elderly men 109/1260, cycling women 131/1040, post-menopausal women 102/1420, and pregnant women 3980/5850.

scholarly journals ERYTHROPOIESIS DURING AMPHIBIAN METAMORPHOSIS

1971 ◽  
Vol 49 (2) ◽  
pp. 390-404 ◽  
Author(s):  
George M. Maniatis ◽  
Vernon M. Ingram
Keyword(s):  
Experimental Error ◽  
Single Cells ◽  
Deae Cellulose ◽  
Fluorescein Isothiocyanate ◽  
Erythroid Cells ◽  
Fluorescent Staining ◽  
Ammonium Sulfate Precipitation ◽  
Blood Smears ◽  
Tetramethylrhodamine Isothiocyanate ◽  
Cellulose Column

Rabbit antibodies specific for the major tadpole and frog hemoglobin components of R. catesbeiana were used for the detection of the two hemoglobins inside single cells. The antisera, after fractionation by ammonium sulfate precipitation and diethylaminoethyl (DEAE)-cellulose chromatography, were conjugated with fluorescein isothiocyanate for the antifrog hemoglobin serum and tetramethylrhodamine isothiocyanate for the antitadpole hemoglobin serum. The conjugated fractions, refractionated by stepwise elution from a DEAE-cellulose column, were used for the fluorescent staining of blood smears, liver tissue imprints, and smears of liver cell suspensions. Both simultaneous and sequential staining with the two fluorescent preparations indicated that larval and adult hemoglobins were not present within the same erythrocyte during metamorphosis. In other experiments, erythroid cells from animals in metamorphosis were spread on agar containing specific antiserum. Precipitates were formed around the cells which contain the particular hemoglobin. The percentages of cells containing either tadpole or frog hemoglobin were estimated within the experimental error of the method. The data showed that the two hemoglobins are in different cells. It is concluded that the hemoglobin change observed during the metamorphosis of R. catesbeiana is due to the appearance of a new population of erythroid cells containing exclusively frog hemoglobin.

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