scholarly journals LOCALIZATION OF CARBONIC ANHYDRASE IN AVIAN GASTRIC MUCOSA, SHELL GLAND AND BONE BY IMMUNOHISTOCHEMISTRY

1974 ◽  
Vol 22 (8) ◽  
pp. 819-825 ◽  
Author(s):  
CAROL V. GAY ◽  
HARALD SCHRAER ◽  
EDWARD J. FALESKI ◽  
ROSEMARY SCHRAER
Keyword(s):  
Carbonic Anhydrase ◽  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Shell Gland ◽  
Significant Finding ◽  
Specific Fluorescence ◽  
Tubular Gland ◽  
Γ Globulins ◽  
Mucosal Lining ◽  
Avian Erythrocyte

Antisera to purified avian erythrocyte carbonic anhydrase (CA) were produced in rabbits and the γ-globulin fractions were isolated and purified by ammonium sulfate precipitation and diethylaminoethyl-Sephadex chromatography. The anti-CA immuno-γ-globulins were tested by immunoelectrophoresis and judged to be highly specific for CA. Fluorescein isothiocyanate-conjugated goat antirabbit γ-globulins were used in the indirect fluorescent antibody localization of CA in cryostat sections. Specific fluorescence of CA was observed in gastric mucosal lining cells, shell gland columnar and tubular gland cells, red blood cells, erythroblasts and osteoclasts. Specific fluorescence was absent in the several immunologic controls, including the blocking test. Specific fluorescence was also lacking in tissue constituents which contain no detectable amounts of CA, i.e., muscle, connective tissue, blood vessels and nuclei. A significant finding in this study is the occurrence of cross-reactions between antibodies to red blood cell CA and CA from other tissues, indicating the immunologic similarity of CA from different tissue sources.

scholarly journals STUDIES ON FLUORESCENT ANTIBODY STAINING

1961 ◽  
Vol 114 (1) ◽  
pp. 89-110 ◽  
Author(s):  
Gerald Goldstein ◽  
Irene S. Slizys ◽  
Merrill W. Chase
Keyword(s):  
Fluorescent Antibody ◽  
Deae Cellulose ◽  
Fluorescein Isothiocyanate ◽  
Gradient Elution ◽  
Fluorescent Staining ◽  
Coupling Ratio ◽  
Specific Fluorescence ◽  
Antibody Staining ◽  
Fluorescent Products ◽  
Elution Chromatography

1. A study has been made of the non-specific fluorescent staining of splenic imprints treated with fluorescent sheep antibody globulins. 2. In tissue imprints made with the spleens of antigen-stimulated animals, no morphological distinction was evident between areas showing non-specific fluorescence and specific fluorescence. 3. Elimination of non-specific fluorescence was not achieved by any one, or any combination of the following: (a) conjugating only gamma globulins with fluorescein isothiocyanate; (b) removal of dialyzable fluorescent products on sephadex, followed by concentration through the use of pressure dialysis; (c) use of crystalline preparations of fluorescein isothiocyanate. 4. Individual preparations of fluorescent antibodies were separated by gradient elution chromatography on diethylaminoethyl (DEAE) cellulose into fractions possessing different numbers of fluorescein radicals per molecule of globulin. 5. The coupling ratio of 50 mg fluorescein isothiocyanate (FITC) per gm of protein, as commonly advocated, can not be recommended for the precise localization of antibody globulin in tissues owing to the capacity of the coupled products to give non-specific fluorescent staining. When crystalline preparations of FITC are used instead of the amorphous product at 50 mg/gm protein, far too high non-specific fluorescence results. 6. A fraction with bright specific fluorescence and no or negligible nonspecific fluorescence was obtained from each fluorescent antibody that was prepared by using 6 to 8 mg of crystalline fluorescein isothiocyanate per gm of globulin and was then subjected to DEAE-cellulose chromatography and gradient elution to eliminate the most highly coupled molecules.

scholarly journals Detection and Quantitation of Minor Erythrocyte Populations in an Admixture by Means of Fluorescent Antibody: Rhesus Monkey

Blood ◽  
1966 ◽  
Vol 28 (4) ◽  
pp. 573-580 ◽  
Author(s):  
ARTHUR E. BOGDEN ◽  
JAMES H. GRAY ◽  
Rosemary Najemy ◽  
Sheryl Giofreda
Keyword(s):  
Rhesus Monkey ◽  
Rapid Method ◽  
Cell Population ◽  
Fluorescent Antibody ◽  
Indirect Fluorescent Antibody ◽  
Specific Fluorescence

Abstract The five rabbit antirhesus monkey erythrocyte specificities of Owen and Anderson1 have been confirmed. An additional specificity, designated "F," has been found in 91.2 per cent of rhesus animals tested. Antiserum reagents with the A and B reactivities have been used in the indirect fluorescent antibody technic to detect and quantitate minor rhesus erythrocyte populations in admixture with a major population. Specific fluorescence was achieved with wet, unfixed preparations and was "all-or-none." A rapid method for the estimation of minor to major cell population ratios well above 1:1000 is presented.

scholarly journals Detection of UV-Induced Thymine Dimers in Individual Cryptosporidium parvum and Cryptosporidium hominis Oocysts by Immunofluorescence Microscopy

2006 ◽  
Vol 73 (3) ◽  
pp. 947-955 ◽  
Author(s):  
B. H. Al-Adhami ◽  
R. A. B. Nichols ◽  
J. R. Kusel ◽  
J. O'Grady ◽  
H. V. Smith
Keyword(s):  
Cryptosporidium Parvum ◽  
Immunofluorescence Microscopy ◽  
Uv Light ◽  
Fluorescein Isothiocyanate ◽  
Immunofluorescence Assay ◽  
Cryptosporidium Hominis ◽  
Specific Fluorescence ◽  
Thymine Dimers ◽  
Texas Red

ABSTRACT To investigate the effect of UV light on Cryptosporidium parvum and Cryptosporidium hominis oocysts in vitro, we exposed intact oocysts to 4-, 10-, 20-, and 40-mJ�cm−2 doses of UV irradiation. Thymine dimers were detected by immunofluorescence microscopy using a monoclonal antibody against cyclobutyl thymine dimers (anti-TDmAb). Dimer-specific fluorescence within sporozoite nuclei was confirmed by colocalization with the nuclear fluorogen 4′,6′-diamidino-2-phenylindole (DAPI). Oocyst walls were visualized using either commercial fluorescein isothiocyanate-labeled anti-Cryptosporidium oocyst antibodies (FITC-CmAb) or Texas Red-labeled anti-Cryptosporidium oocyst antibodies (TR-CmAb). The use of FITC-CmAb interfered with TD detection at doses below 40 mJ�cm−2. With the combination of anti-TDmAb, TR-CmAb, and DAPI, dimer-specific fluorescence was detected in sporozoite nuclei within oocysts exposed to 10 to 40 mJ�cm−2 of UV light. Similar results were obtained with C. hominis. C. parvum oocysts exposed to 10 to 40 mJ�cm−2 of UV light failed to infect neonatal mice, confirming that results of our anti-TD immunofluorescence assay paralleled the outcomes of our neonatal mouse infectivity assay. These results suggest that our immunofluorescence assay is suitable for detecting DNA damage in C. parvum and C. hominis oocysts induced following exposure to UV light.

Effect of Reserpine Pretreatment on Avian Erythrocyte Carbonic Anhydrase Activation by Isoproterenol

Pharmacology ◽  
1994 ◽  
Vol 49 (2) ◽  
pp. 112-120 ◽  
Author(s):  
Immaculata N.A. Igbo ◽  
Charles E. Reigel, Jr. ◽  
Ingrid M. Greene ◽  
Alexander D. Kenny
Keyword(s):  
Carbonic Anhydrase ◽  
Avian Erythrocyte ◽  
Erythrocyte Carbonic Anhydrase

THE CYTOLOGICAL LOCALIZATION OF HUMAN CHORIONIC GONADOTROPIN

1963 ◽  
Vol 41 (12) ◽  
pp. 2517-2521 ◽  
Author(s):  
Arthur Leznoff ◽  
Bernard A. Davis
Keyword(s):  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Fluorescent Antibody ◽  
Placental Tissue ◽  
Giant Cells ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Chorionic Villi ◽  
Specific Fluorescence ◽  
Cytological Localization

The indirect fluorescent antibody technique was used to determine the cellular site of human chorionic gonadotropin (H.C.G.) in normal and toxaemic placentas, and in choriocarcinomas. In placental tissue specific fluorescence was located in the syncytial cells of the chorionic villi but not in the cytotrophoblast cells. In choriocarcinomas specific fluorescence was seen in the syncytial giant cells. No distinct difference could be demonstrated between normal and "toxic" placentas. Differences in the content of H.C.G. in placentas at various stages of pregnancy were noted. Maximum amounts were demonstrated in tissue of less than 14 weeks gestation. Lesser quantities could be seen in more mature placentas and some specific fluorescence could be seen in most full term placentas.

Comparison of detection specificity of nitrifying bacteria in biofilm using fluorescence in situ hybridization and in situ fluorescent antibody methods

2003 ◽  
Vol 47 (5) ◽  
pp. 129-132
Author(s):  
N. Noda ◽  
Y. Ebie ◽  
M. Matsumura ◽  
S. Tsuneda ◽  
A. Hirata ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Specific Fluorescence ◽  
Antibody Method ◽  
In Situ Detection ◽  
Detection Specificity ◽  
Fluorescent Antibody Method

The in situ fluorescent antibody and fluorescence in situ hybridization (FISH) methods are very useful in the in situ detection of specific bacteria like nitrifiers in a biofilm. In this study, simultaneous staining using the FISH and in situ fluorescent antibody methods was examined. As a result, no specific fluorescence was observed with either method when FISH was performed followed by the in situ fluorescent antibody method; however, when the in situ fluorescent antibody method was performed first followed by FISH, specific fluorescence was observed in both cases. Moreover, it was suggested that the detection specificities of FISH and the in situ fluorescent antibody method are almost identical.

Enterotoxin formation by Clostridium perfringens type A studied by the use of fluorescent antibody

1977 ◽  
Vol 23 (7) ◽  
pp. 908-915 ◽  
Author(s):  
L. Niilo
Keyword(s):  
Clostridium Perfringens ◽  
Fluorescent Antibody ◽  
Mature Spore ◽  
Fluorescein Isothiocyanate ◽  
Type A ◽  
Entire Length

Fluorescein isothiocyanate-conjugated antibody to purified enterotoxin of Clostridium perfringens was used to study the intracellular formation of enterotoxin by this organism. Enterotoxin was detected at 4 h of growth at the end of the cell containing forespore. With the development of the spore, enterotoxin accumulation continued and involved the entire length of the cell until its lysis with the release of enterotoxin and mature spore. The spores did not contain demonstrable enterotoxin. Only a certain number of the sporulated cells of the enterotoxigenic strains studied produced this toxin. The amount of enterotoxin produced varied with sporulation percentage, and between strains and individual cells.

scholarly journals Description of a polyvalent conjugate and a new serogroup of Bacteroides melaninogenicus by fluorescent antibody staining

1976 ◽  
Vol 3 (5) ◽  
pp. 506-512
Author(s):  
D W Lambe ◽  
R C Jerris
Keyword(s):  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Rapid Identification ◽  
Antibody Staining ◽  
Biochemical Testing ◽  
Fluorescent Antibody Staining

A polyvalent conjugate (fluorescein isothiocyanate-labeled antibody reagent) containing serogroups A, B, and C conjugates was prepared. This polyvalent conjugate gave a positive fluorescent antibody (FA) stain with 49 stains of Bacteroides melaninogenicus representing serogroups A, B, and C. When additional strains (92 strains) of the three subspecies of B. melaninogenicus were examined by the FA stain, with A, B, and C, and polyvalent conjugates, nine strains of B. melaninogenicus subsp. intermedius failed to give a positive stain with any conjugate. Therefore, an FA conjugate was prepared with the antiserum to one of these strains (532-70A); all nine strains stained positively with this conjugate. These nine strains were biochemically characteristic of B. melaninogenicus subsp. intermedius; thus, these strains were designated as a new serogroup, serogroup C-1. A new polyvalent conjugate containing serogroups A, B, C, and C-1 was prepared. This polyvalent conjugate stained positively with 23 representative strains from serogroups A, B, C, and C-1. The new conjugates failed to stain positively with other anaerobes and aerobes tested. The four individual conjugates, as well as the polyvalent conjugate, may be used for a more rapid identification of B. melaninogenicus than is possible by biochemical testing.

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