Localization of H-2Kk in developing mouse palates using monoclonal antibody

Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 45-60
Author(s):  
Melnick Melnick ◽  
Tina Jaskoll ◽  
Mary Marazita
Keyword(s):  
Monoclonal Antibody ◽  
Acetic Acid ◽  
Monoclonal Antibodies ◽  
Glacial Acetic Acid ◽  
Corticosteroid Treatment ◽  
Basement Membranes ◽  
Spatiotemporal Distribution ◽  
Embryonic Tissue ◽  
Epithelial Fusion

Using monoclonal antibodies to H-2Kk antigen, we sought to develop a reproduceable method of in situ localization in embryonic tissue and to determine whether there are specific patterns of H-2 localization in time and space in the developing palatal tissues of B10.A(H-2a) embryonic mice, with and without corticosteroid pretreatment at 12 days gestation. Our procedure employs ethanol-glacial acetic acid fixation, paraplast embedding, and enzymatic predigestion with purified hyaluronidase and neuraminidase. H-2 antigens were detected in palatal mesenchyme as well as basement membranes but not in oral or nasal epithelium. The pattern of distribution in mesenchyme of untreated embryos changed with progressive shelf development: vertical → horizontal → epithelial fusion → epithelial seam degeneration → mesenchymal confluence. Although the palatal shelves of treated embryos remained vertical, corticosteroid treatment does not appear to alter the detectable spatiotemporal distribution of H-2 antigens in developing palates of embryonic B10.A mice.

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.

scholarly journals Monoclonal antibodies to laminin reveal the heterogeneity of basement membranes in the developing and adult mouse tissues.

1984 ◽  
Vol 98 (3) ◽  
pp. 971-979 ◽  
Author(s):  
Y J Wan ◽  
T C Wu ◽  
A E Chung ◽  
I Damjanov
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Adult Mouse ◽  
Basement Membranes ◽  
Newborn Mouse ◽  
Preimplantation Stage ◽  
Mouse Tissues ◽  
Reichert's Membrane ◽  
Anatomic Locations ◽  
Immunohistochemical Studies

Two monoclonal antibodies raised against laminin isolated from a mouse parietal yolk sac cell line were used for immunohistochemical studies of basement membranes of the mouse embryo and various fetal and adult tissues. No immunoreactivity with either of the two monoclonal antibodies could be detected in the preimplantation-stage embryos, although it has been shown that these embryos contain extracellular laminin reactive with the conventional polyclonal antilaminin antibodies. Reichert's membrane in early postimplantation stages of development reacted with the monoclonal antibody LAM-I but not with the antibody LAM-II. However, from day 8 of pregnancy onward the Reichert's membrane reacted with both antibodies. Basement membranes of the embryo proper were unreactive with both monoclonal antibodies until day 12 of pregnancy. By day 14 some basement membranes of the fetal tissues became reactive with one or both monoclonal antibodies, whereas others remained still unreactive. In the 17-d fetus and the newborn mouse most of the basement membranes reacted with both monoclonal antibodies, whereas others still reacted with only one. Similar heterogeneity in the immunoreactivity of basement membranes of various tissues was noted in the adult mouse as well. These results indicate that the immunoreactivity of laminin in the extracellular matrix changes during development and that the basement membranes in various anatomic locations display heterogeneity even in the adult mouse.

scholarly journals Synthesis of Bioactive Fluorinated 10H-Phenothiazines and their Sulfone Derivatives

2008 ◽  
Vol 5 (s1) ◽  
pp. 1063-1068 ◽  
Author(s):  
Yogesh Dixit ◽  
Rahul Dixit ◽  
Naveen Gautam ◽  
D. C. Gautam
Keyword(s):  
Hydrogen Peroxide ◽  
Acetic Acid ◽  
Spectroscopic Data ◽  
Halogen Atom ◽  
Glacial Acetic Acid ◽  
Present Communication ◽  
Smiles Rearrangement ◽  
Antimicrobial Studies ◽  
Sulfone Derivatives

The present communication deals with the synthesis of a series of fluorinated 10H-phenothiazines. 10H-phenothiazines is prepared by Smiles rearrangement of substituted 2-foramido-2´-nitrodiphenylsulfide. Substituted 2-foramido-2´-nitrodiphenylsulfide were obtained by the reaction of 2-amino-3-fluorobenzenethiol witho-halonitrobenzenes followed by formylation and 1-nitro/1-halo-10H-phenothiazines have been prepared by the reaction of substituted 2-aminobenzenethiols with reactiveo-halonitrobenzene containing a nitro group or halogen atom ato-position to the reactive halogen atom directly yielded 1-nitro/1-halo-10H-phenothiazines in situ. 10H-phenothiazine sulfone derivatives have been synthesized by the oxidation of 10H-phenothiazines by 30% hydrogen peroxide in glacial acetic acid. The structure of the synthesized compounds has been characterized by spectroscopic data and elemental analysis. Antimicrobial studies of the synthesized compounds have also been included.

Response Surface Methodology for Optimization of Epoxidized Trimethylolpropane Ester Synthesis from Palm Oil

2011 ◽  
Vol 9 (1) ◽  
Author(s):  
Ferra Naidir ◽  
Robiah Yunus ◽  
Irmawati Ramli ◽  
Tinia I. Mohd. Ghazi
Keyword(s):  
Hydrogen Peroxide ◽  
Response Surface Methodology ◽  
Acetic Acid ◽  
Regression Model ◽  
Palm Oil ◽  
Iodine Value ◽  
Glacial Acetic Acid ◽  
Oxirane Oxygen ◽  
Dependent Variables

To improve the oxidative stability of the palm oil-based biolubricant, the fatty acid double bonds in palm oil-based trimethylolpropane ester (TMP ester) was converted into an oxirane ring via an in-situ epoxidation method. The epoxidized TMP ester was produced from a reaction between TMP ester and peracetic acid which was prepared in-situ by reacting glacial acetic acid with hydrogen peroxide in the presence of concentrated sulphuric acid. The response surface methodology was applied using a central composite design technique to optimize the conditions of the epoxidation reaction to produce the epoxidized TMP ester. The effects of four independent variables namely concentration of acetic acid (0-2 mol), concentration of hydrogen peroxide (1.5-9.5 mol), temperature of reaction (30-110°C) and reaction time (0.5-26.5 h) on the three dependent variables; percentage of oxirane oxygen, iodine value, and hydroxyl value were studied. A second-order polynomial multiple regression model was employed to predict the three dependent variables under optimum conditions of 0.59 mol of glacial acetic acid, 7.5 mol of hydrogen peroxide concentration, at temperature of 50°C and reaction times of 7 h. The optimum values of percentage of oxirane oxygen, iodine value, and hydroxyl value were 4.01%, 1.94%, and 0.43% respectively. The analysis of variance yielded a high coefficient of determination value of 0.9395-0.9880, hence indicating the fitness of the second-order regression model to the experimental data.

scholarly journals Generation of monoclonal antibodies detecting specific epitopes in locust antennae

1993 ◽  
Vol 175 (1) ◽  
pp. 45-57
Author(s):  
J. Strotmann ◽  
I. Boekhoff ◽  
S. Goggerle ◽  
H. Breer
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Protein Synthesis ◽  
Monoclonal Antibodies ◽  
Embryonic Development ◽  
High Molecular Weight ◽  
Time Course ◽  
Antigen Expression

1. Following a tissue-specific screening paradigm, monoclonal antibodies have been generated that interact with distinct subpopulations of cells in locust antennae. 2. Antigens were identified as high molecular weight components. 3. Immunoreactivity was not detectable during embryonic development, but rapidly appeared within a few hours of hatching. 4. The time course of antigen expression in antennal cells could be followed in situ as well as in vitro. 5. Expression of monoclonal antibody B14/6D2-like immunoreactivity was prevented by blocking protein synthesis with cycloheximide.

scholarly journals An Investigation of the Antigenic Characteristics of the Renal Glomerulus in the Rat

2021 ◽  
Author(s):  
◽  
Maurice James Nicol
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Animal Models ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Immune Complexes ◽  
Capillary Wall ◽  
Glomerular Capillary Wall ◽  
Glomerular Capillary

<p>The finding of a granular deposition of immunoglobulin in the kidney in experimental animal models of glomerulonephritis has been been interpreted as resulting from the random deposition of immune complexes in the glomeruli. Recent data suggests that although immune complex deposition may be an important factor in some forms of glomerulonephritis, the in situ formation of immune complexes between circulating anti-kidney antibodies and fixed glomerular capillary wall antigens may also be a significant factor in the pathogenesis of some animal models of glomerulonephritis. To examine the characteristics of discontinuously represented glomerular capillary wall antigens in the rat, monoclonal antibodies were generated against a glomerular plasma membrane fraction, depleted of glomerular basement membrane, prepared from isolated Lewis rat glomeruli. A total of 17 hybridomas, generated from the fusion of splenocytes obtained from mice immunised with the glomerular membrane fraction produced monoclonal antibodies which reacted with discontinuously represented antigens in the glomerulus and renal tubules. One further hybridoma secreted a monoclonal antibody which reacted with an antigen present on glomerular and tubular nuclear membranes. No hybridomas were produced which secreted a monoclonal antibody which reacted with linearly arrayed glomerular basement membrane antigens. Two of these monoclonal antibodies, both of the IgM subclass and code-named PH7 and SC5, produced a heavy granular glomerular staining pattern when examined by indirect immunofluorescence microscopy. Neither monoclonal antibody was kidney specific, with reactivity being demonstrated with a number of non-renal tissues. When administered intravenously to normal Lewis rats both SC5 and PH7 induced a mild proteinuric lesion. The proteinuria was not associated with histopathological changes at the light or electron microscope level. Immunoblotting experiments revealed that SC5 reacted predominantly with a protein band of 96 kDa present in detergent extracts of isolated glomeruli and glomerular plasma membranes. PH7 was shown to react with three low molecular weight proteins of 14, 13 and 11 kDa The findings of this study demonstrate the potential for a nephritogenic response to occur following the in situ formation of immune complexes between circulating anti-kidney antibodies and discontinuously arrayed non-glomerular, basement membrane glomerular capillary wall antigens characterised by granular immunofluorescence patterns,in animal models of glomerulonephritis.</p>

scholarly journals An Investigation of the Antigenic Characteristics of the Renal Glomerulus in the Rat

2021 ◽  
Author(s):  
◽  
Maurice James Nicol
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Animal Models ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Immune Complexes ◽  
Capillary Wall ◽  
Glomerular Capillary Wall ◽  
Glomerular Capillary

<p>The finding of a granular deposition of immunoglobulin in the kidney in experimental animal models of glomerulonephritis has been been interpreted as resulting from the random deposition of immune complexes in the glomeruli. Recent data suggests that although immune complex deposition may be an important factor in some forms of glomerulonephritis, the in situ formation of immune complexes between circulating anti-kidney antibodies and fixed glomerular capillary wall antigens may also be a significant factor in the pathogenesis of some animal models of glomerulonephritis. To examine the characteristics of discontinuously represented glomerular capillary wall antigens in the rat, monoclonal antibodies were generated against a glomerular plasma membrane fraction, depleted of glomerular basement membrane, prepared from isolated Lewis rat glomeruli. A total of 17 hybridomas, generated from the fusion of splenocytes obtained from mice immunised with the glomerular membrane fraction produced monoclonal antibodies which reacted with discontinuously represented antigens in the glomerulus and renal tubules. One further hybridoma secreted a monoclonal antibody which reacted with an antigen present on glomerular and tubular nuclear membranes. No hybridomas were produced which secreted a monoclonal antibody which reacted with linearly arrayed glomerular basement membrane antigens. Two of these monoclonal antibodies, both of the IgM subclass and code-named PH7 and SC5, produced a heavy granular glomerular staining pattern when examined by indirect immunofluorescence microscopy. Neither monoclonal antibody was kidney specific, with reactivity being demonstrated with a number of non-renal tissues. When administered intravenously to normal Lewis rats both SC5 and PH7 induced a mild proteinuric lesion. The proteinuria was not associated with histopathological changes at the light or electron microscope level. Immunoblotting experiments revealed that SC5 reacted predominantly with a protein band of 96 kDa present in detergent extracts of isolated glomeruli and glomerular plasma membranes. PH7 was shown to react with three low molecular weight proteins of 14, 13 and 11 kDa The findings of this study demonstrate the potential for a nephritogenic response to occur following the in situ formation of immune complexes between circulating anti-kidney antibodies and discontinuously arrayed non-glomerular, basement membrane glomerular capillary wall antigens characterised by granular immunofluorescence patterns,in animal models of glomerulonephritis.</p>

A web-like network of type vi collagen in human skin is revealed by immuno-electron microscopy

1988 ◽  
Vol 46 ◽  
pp. 382-383
Author(s):  
Douglas R. Keene ◽  
Robert W. Glanville ◽  
Eva Engvall
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Human Skin ◽  
Basement Membranes ◽  
Primary Antibody ◽  
Fat Cells ◽  
Secondary Antibody ◽  
Type Vi Collagen ◽  
Dermal Matrix ◽  
Immuno Electron Microscopy

A mouse monoclonal antibody (5C6) prepared against human type VI collagen (1) has been used in this study to immunolocalize type VI collagen in human skin. The enbloc method used involves exposing whole tissue pieces to primary antibody and 5 nm gold conjugated secondary antibody before fixation, and has been described in detail elsewhere (2).Biopsies were taken from individuals ranging in age from neonate to 65 years old. By immuno-electron microscopy, type VI collagen is found to be distributed as a fine branching network closely associated with (but not attached to) banded collagen fibrils containing types I and III collagen (Fig. 1). It appears to enwrap fibers, to weave between individual fibrils within a fiber, and to span the distance separating fibers, creating a “web-like network” which entraps fibers within deep papillary and reticular dermal layers (Fig. 2). Relative to that in the dermal matrix, the concentration of type VI collagen is higher around endothelial basement membranes limiting the outer boundaries of nerves, capillaries, and fat cells (Fig. 3).

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