Data on the inhibition of RNase inhibitor activity by a monoclonal antibody as assessed by microfluidics-based RNA electrophoresis

Ehrlich ascites cells were treated with actinomycin D before transplantation. These cells, collected after 7 days, show increased alkaline RNase II, acid RNase II, and phosphodiesterase II activities and low RNase inhibitor activity as compared to untreated tumor cells collected 7 days after transplantation. RNase I, phosphodiesterase I, and acid phosphatase activities were unchanged. Smaller increases in the same enzyme activities, but no change in RNase inhibitor activity, were observed when untreated cells collected after 4 days were compared to untreated cells collected after 7 days of growth. Ehrlich ascites cells collected after 4 days synthesized protein and RNA at a slightly faster rate than those collected after 7 days. Actinomycin D treated cells synthesized protein at a rate identical to that of the control cells; net synthesis of RNA, however, was significantly reduced. One possible reason for this may be the higher RNase and phosphodiesterase activities in actinomycin D treated cells.

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Enzyme-linked immunosorbent assay (ELISA) of factor VIII antigen using a monoclonal antibody devoid of factor VIII inhibitor activity.

Blood & Vessel ◽

10.2491/jjsth1970.18.48 ◽

1987 ◽

Vol 18 (1) ◽

pp. 48-59 ◽

Cited By ~ 2

Author(s):

Midori SHIMA ◽

Akira YOSHIOKA ◽

Ichiro TANAKA ◽

Toshiharu FUJIWARA ◽

Shunsuke IMAI ◽

...

Keyword(s):

Monoclonal Antibody ◽

Factor Viii ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Factor Viii Inhibitor ◽

Inhibitor Activity ◽

Factor Viii Antigen ◽

Viii Inhibitor

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Properties and fluctuations in vivo of rat liver antizyme inhibitor

Biochemical Journal ◽

10.1042/bj2590839 ◽

1989 ◽

Vol 259 (3) ◽

pp. 839-845 ◽

Cited By ~ 24

Author(s):

Y Murakami ◽

S Matsufuji ◽

M Nishiyama ◽

S Hayashi

Keyword(s):

Monoclonal Antibody ◽

Affinity Chromatography ◽

Rat Liver ◽

Regulatory Protein ◽

Inhibitor Activity ◽

Reversible Binding ◽

Structural Resemblance ◽

Antizyme Inhibitor ◽

Tissue Extracts

Antizyme inhibitor was highly purified from rat liver by using affinity chromatography. It has some structural resemblance to ornithine decarboxylase (ODC), as judged from Mr, immunoreactivity and reversible binding with antizyme. However, unlike hepatic amounts of ODC and ODC-antizyme complex, that of antizyme inhibitor did not show much fluctuation upon putrescine treatment, whereas it decreased as rapidly as ODC decay in the presence of cycloheximide. These results suggested that antizyme inhibitor is an independent regulatory protein rather than a derivative of ODC. Changes in hepatic amounts of antizyme inhibitor, antizyme and ODC upon feeding suggested that antizyme inhibitor may play a role in ODC regulation by trapping antizyme and thereby suppressing ODC degradation. A monoclonal antibody to rat liver antizyme inhibitor was obtained. This antibody was shown to be utilizable for a simple assay of antizyme-inhibitor activity in tissue extracts.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100103978 ◽

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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High-voltage electron microscopy of subretinal scar formation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100122836 ◽

1992 ◽

Vol 50 (1) ◽

pp. 486-487

Author(s):

G.E. Korte ◽

M. Marko ◽

G. Hageman

Keyword(s):

Electron Microscopy ◽

Monoclonal Antibody ◽

Retinal Pigment Epithelium ◽

Glial Cells ◽

High Voltage ◽

Pigment Epithelium ◽

Retinal Pigment ◽

Sodium Iodate ◽

High Voltage Electron Microscopy ◽

Voltage Electron

Sodium iodate iv. damages the retinal pigment epithelium (RPE) in rabbits. Where RPE does not regenerate (e.g., 1,2) Muller glial cells (MC) forma subretinal scar that replaces RPE. The MC response was studied by HVEM in 3D computer reconstructions of serial thick sections, made using the STEREC0N program (3), and the HVEM at the NYS Dept. of Health in Albany, NY. Tissue was processed for HVEM or immunofluorescence localization of a monoclonal antibody recognizing MG microvilli (4).

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Identification of the pulmonary syndrome hantavirus by direct and colloidal gold immune Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100169109 ◽

1994 ◽

Vol 52 ◽

pp. 274-275

Author(s):

C. D. Humphrey ◽

C.S. Goldsmith ◽

L. Elliott ◽

S.R. Zaki

Keyword(s):

Electron Microscopy ◽

United States ◽

Monoclonal Antibody ◽

Colloidal Gold ◽

Phosphotungstic Acid ◽

Uranyl Acetate ◽

Hantavirus Pulmonary Syndrome ◽

Deer Mice ◽

Immune Electron Microscopy ◽

Negative Stain

An outbreak of unexplained acute pulmonary syndrome with high fatality was recognized in the spring of 1993 in the southwestern United States. The cause of the illness was quickly identified serologically and genetically as a hantavirus and the disease was named hantavirus pulmonary syndrome (HPS). Recently, the virus was isolated from deer mice which had been trapped near the homes of HPS patients, and cultivated in Vero E6 cells. We identified the cultivated virus by negative-stain direct and colloidal gold immune electron microscopy (EM).Virus was extracted, clarified, and concentrated from unfixed and 0.25% glutaraldehyde fixed supernatant fluids of infected Vero E6 cells by a procedure described previously. Concentrated virus suspensions tested by direct EM were applied to glow-discharge treated formvar-carbon filmed grids, blotted, and stained with 0.5% uranyl acetate (UA) or with 2% phosphotungstic acid (PTA) pH 6.5. Virus suspensions for immune colloidal gold identification were adsorbed similarly to filmed grids but incubated for 1 hr on drops of 1:50 diluted monoclonal antibody to Prospect Hill virus nucleoprotein or with 1:50 diluted sera from HPS virus infected deer mice.

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