scholarly journals LOCALIZATION OF MYOGLOBIN IN HUMAN SKELETAL MUSCLE USING FLUORESCENT ANTIBODY TECHNIQUE

1967 ◽  
Vol 15 (8) ◽  
pp. 436-441 ◽  
Author(s):  
LAWRENCE J. KAGEN ◽  
RAYA GUREVICH
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Fluorescent Antibody ◽  
Intracellular Localization ◽  
Rabbit Antiserum ◽  
Rabbit Serum ◽  
Fluorescent Antibody Technique ◽  
Normal Rabbit Serum ◽  
Antibody Technique ◽  
Human Myoglobin

Rabbit antiserum to human myoglobin was used with the indirect fluorescent antibody technique to localize this protein in human skeletal muscle. Specific fluorescence was noted, in rapidly frozen and acetone-fixed sections, to be located at the transverse striations, at the sarcolemmal regions and at certain fibrillar structures within the cell. The antibody fluorescence reaction was shown to be specific for myoglobin, and was not produced by normal rabbit serum of serum of rabbits immunized with bacterial antigens. The reaction was abolished by prior absorption of the antimyoglobin serum with myoglobin, and was found to be absent in tissues deficient in myoglobin (lung, kidney, spleen, liver and uterus). Omission of acetone fixation or delayed freezing resulted in leakage of myoglobin from the cell and loss of specific intracellular localization. Sarcolemmal localization appeared to be somewhat more stable.

scholarly journals LOCALIZATION OF ADRENOCORTICOTROPIC HORMONE BY HISTOCHEMICAL AND IMMUNOCHEMICAL METHODS

1951 ◽  
Vol 94 (1) ◽  
pp. 21-30 ◽  
Author(s):  
John M. Marshall
Keyword(s):  
Adrenocorticotropic Hormone ◽  
Fluorescent Antibody ◽  
Rabbit Serum ◽  
Fluorescent Antibody Technique ◽  
Normal Rabbit Serum ◽  
Protein Antigens ◽  
Antibody Technique ◽  
Selective Staining ◽  
Highly Active ◽  
Hog Kidney

The fluorescent antibody technique was adapted to the localization of native protein antigens in cells and tissues. This method was applied specifically to the localization of adrenocorticotropic hormone in the pituitary gland. An antiserum to hog ACTH was produced in an adrenalectomized rabbit. The γ2-globulin fraction of the serum was conjugated with fluorescein. After purification, the fluorescent antibody solution stained selectively the cytoplasm of basophil cells of the hog pituitary. No cells of sheep or beef pituitary or of hog kidney were stained. A fluorescent globulin solution prepared from normal rabbit serum gave no selective staining in any of these tissues. Immunochemical tests showed that the fluorescent antibody gave a precipitin reaction with a highly active ACTH preparation of low molecular weight. The supernatant solution from this reaction showed a loss of hormone activity.

scholarly journals DEMONSTRATION OF INTRACELLULAR CARRAGEENAN BY A FLUORESCENT ANTIBODY TECHNIQUE IN GRANULOMA TISSUE

1972 ◽  
Vol 20 (12) ◽  
pp. 991-994 ◽  
Author(s):  
SUZANNE M. RICHER ◽  
ESTHER L. MCCANDLESS
Keyword(s):  
Fluorescent Antibody ◽  
Intracellular Localization ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Staining ◽  
Fluorescent Antibody Staining ◽  
Granuloma Tissue

Intracellular localization of λ-carrageenan in the carrageenan granuloma has been demonstrated by a specific fluorescent antibody to the polymer. In general, the fluorescent antibody staining correlated with the intracellular metachromatic staining. The fluorescent antibody technique did not confirm that the extracellular metachromasia was due to λ-carrageenan.

scholarly journals INTRACELLULAR LOCALIZATION OF TYPE 4 ADENOVIRUS

1959 ◽  
Vol 109 (1) ◽  
pp. 85-96 ◽  
Author(s):  
Georgiana S. Boyer ◽  
Floyd W. Denny ◽  
Harold S. Ginsberg
Keyword(s):  
Structural Changes ◽  
Infectious Virus ◽  
Fluorescent Antibody ◽  
Intracellular Localization ◽  
Adenovirus Type ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Nuclear Changes ◽  
Infected Cells ◽  
Intracellular Antigen

HeLa cell cultures infected with adenovirus type 4 were studied by light and phase-contrast microscopy and by the fluorescent antibody technique for visualization of intracellular antigen. The findings were correlated with the growth curve of infectious virus, determined from companion cultures. The results indicated that those cells undergoing characteristic structural changes observable by light microscopy were those which contain viral antigen. The distribution of the majority of the antigen within the infected cells corresponded to that of the regularly aligned granules and crystal-like masses seen in the nuclei of cells in stained and in unfixed cultures. The production of infectious virus was closely correlated with the development of the characteristic nuclear changes.

scholarly journals Localization of creatine kinase isoenzymes in myofibrils. I. Chicken skeletal muscle.

1977 ◽  
Vol 75 (2) ◽  
pp. 297-317 ◽  
Author(s):  
T Wallimann ◽  
D C Turner ◽  
H M Eppenberger
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Developmental Stages ◽  
Fluorescent Antibody ◽  
Dry Weight ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Peptide Pattern ◽  
Integral Element ◽  
Low Ionic Strength

Purified, repeatedly washed, skeletal muscle myofibrils contain approx. 0.2 U of creatine kinase (CK) activity (equivalent to 2.5 micrograms CK) per milligram dry weight; this firmly bound CK activity is estimated to represent 3-5% of the total cellular CK. It had been shown previously that the myofibrillar CK, which can be quantitatively extracted at low ionic strength and purified to homogeneity, is very similar, if not identical, to the bulk MM-CK. It is shown that the two protein preparations also have the same peptide pattern after cyanogen bromide fractionation and very similar specific activities, confirming their identity. The earlier demonstration that the bound CK is specifically located at the M-lines of isolated myofibrils has been confirmed by immunofluorescence. Antibodies directed against purified MM- and BB-CK were used in the indirect fluorescent antibody technique to study the specificity of myofibril binding sites for different forms of CK. With myofibrils from adult muscle, which has only MM-CK, as well as from early developmental stages in which BB-CK is the predominant isoenzyme, M-type CK was localized exclusively at the M-line, while greater or lesser amounts of B-type CK were found at the Z-line. The data provide strong evidence that the MM-CK at the M-lines in skeletal myofibrils is not adventitiously bound but is rather an integral element in the M-line structure. The amount of CK bound is reasonably consistent with the earlier proposal that the CK molecules might be the transverse M-bridges and appears to be sufficient to regenerate all of the ATP hydrolyzed during muscle contraction.

scholarly journals Detection of Edwardsiella tardain catfish (Clarias sp.)by fluorescent antibody technique (FAT)

2013 ◽  
Vol 11 (1) ◽  
pp. 96
Author(s):  
. Firma ◽  
Rahman Rizky Amalia ◽  
Utami Sari ◽  
Chotimah Chusbul ◽  
Abdulgani Amri Siregar
Keyword(s):  
Fluorescent Antibody ◽  
Culture Method ◽  
Infected Fish ◽  
Edwardsiella Tarda ◽  
Rabbit Serum ◽  
Fluorescent Antibody Technique ◽  
Fish Disease ◽  
Antibody Technique ◽  
Specificity Test ◽  
Kidney Liver

The fluorescent antibody technique (FAT) can be used to detectEdwardsiellatarda rapidly. As prelimenary step, it have been performed purity test of bacteria by PCR, specificity test of mouse anti-E. tarda monoclonal antibody by blocking using chicken and rabbit serum, and optimization of conjugate secondary antibody mouse IgG-H&amp;L (FITC) dillution rate. Fourty eights catfish were intraperitoneally injected by 0,3 mL of E. tarda with different concentration, namely: 102 CFU/mL, 103 CFU/mL, 105 CFU/mL. Kidney, spleen, and liver from three fishes in each treatment were collected at interval time of six hours, 12 hours, 24 hours, 48 hours after infection. The results showed that E. tarda could be detected in fish infected with 102 CFU of E. tarda after six hours of injection in kidney,liver, and spleen of infected fish. Hence, FAT is faster than detection by bacterial culture method, and this technique can be useful to prevent the spread of fish disease.<br /> <br />Keywords: fluorescent antibody technique, Edwardsiella tarda, detection, catfish<br /><br />

Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

Rubella Antibodies in Human Serum: Detection by the Indirect Fluorescent-Antibody Technique

Science ◽  
1964 ◽  
Vol 145 (3635) ◽  
pp. 943-945 ◽  
Author(s):  
G. C. Brown ◽  
H. F. Maassab ◽  
J. A. Veronelli ◽  
T. J. Francis
Keyword(s):  
Human Serum ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Indirect Fluorescent Antibody ◽  
Antibody Technique ◽  
Serum Detection

scholarly journals LOCALIZATION OF UROMUCOID IN HUMAN KIDNEY AND IN SECTIONS OF HUMAN KIDNEY STONE WITH THE FLUORESCENT ANTIBODY TECHNIQUE

1965 ◽  
Vol 13 (3) ◽  
pp. 155-160 ◽  
Author(s):  
H. J. KEUTEL
Keyword(s):  
Kidney Stones ◽  
Kidney Stone ◽  
Fluorescent Antibody ◽  
Human Kidney ◽  
Fluorescent Antibody Technique ◽  
Renal Tubules ◽  
Antibody Technique ◽  
The Matrix ◽  
Normal Human ◽  
The Common

Fluorescent labeled antibodies were used for the demonstration of uromucoid. This urine specific mucoprotein is demonstrably present only in the epithelial cells of the proximal segments of the normal human renal tubules and in the matrix of human kidney stones of all the common crystalline compositions.

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