scholarly journals One-step double immunolabeling of mouse interdigitating reticular cells: simultaneous application of pre-formed complexes of monoclonal rat antibody M1-8 with horseradish peroxidase-linked anti-rat immunoglobulins and of monoclonal mouse anti-Ia antibody with alkaline phosphatase-coupled anti-mouse immunoglobulins.

1991 ◽  
Vol 39 (12) ◽  
pp. 1719-1723 ◽  
Author(s):  
T Krenács ◽  
H Uda ◽  
S Tanaka
Keyword(s):  
Alkaline Phosphatase ◽  
Horseradish Peroxidase ◽  
B Lymphocytes ◽  
Enzyme Reaction ◽  
Molecular Complexes ◽  
Simultaneous Application ◽  
One Step ◽  
Reticular Cells ◽  
Secondary Antibodies ◽  
Double Immunolabeling

A novel one-step double immunolabeling method was elaborated on the basis of the simultaneous application of preformed molecular complexes of two primary antibodies with their specific secondary antibodies labeled with different enzymes. Treatment with a rat monoclonal antibody (MAb), M1-8, pre-coupled with horseradish peroxidase-linked sheep anti-rat immunoglobulins, and enzyme reaction revealed by the 3-amino-9-ethylcarbazole/hydrogen peroxide reaction, resulted in red-brown intracytoplasmic staining of interdigitating reticular cells in the lymph nodes of Balb/c mice. Another molecular complex, made of mouse anti-Ia MAb with alkaline phosphatase-linked rabbit anti-mouse immunoglobulins, applied at the same time and then developed with naphthol AS-BI-phosphate/fast blue BB as substrate, yielded blue surface staining of this cell type in addition to labeling of B-lymphocytes. The method described provides the possibility of relatively rapid double antigen detection where the binding sites of the secondary antibodies are saturated by the specific primary immunoglobulins. This approach seems to avoid nonspecific binding of primary antibodies to Fc receptors, and the unwanted binding of secondary antibodies with cell surface immunoglobulins on B-lymphocytes or with crossreactive primary antibodies used in the other sequence, if the primary antibodies and the tissue are the same or crossreactive animal species.

scholarly journals Assessment of a method for immunochemical detection of antigen on nitrocellulose membranes.

1989 ◽  
Vol 37 (2) ◽  
pp. 257-263 ◽  
Author(s):  
N M Chu ◽  
A J Janckila ◽  
J H Wallace ◽  
L T Yam
Keyword(s):  
Alkaline Phosphatase ◽  
Horseradish Peroxidase ◽  
Reaction Medium ◽  
Intestinal Alkaline Phosphatase ◽  
Dot Blot ◽  
Western Blots ◽  
Calf Intestinal Alkaline Phosphatase ◽  
Immunochemical Detection ◽  
Staining Methods ◽  
Secondary Antibodies

Immunoblotting techniques are widely used for detection of antigen immobilized on nitrocellulose membranes. There are many immunolabeling methods and staining methods available to disclose the presence of antigen in such techniques. Five common staining methods each for alkaline phosphatase and horseradish peroxidase were examined. The staining methods with the highest sensitivity and the lowest background were selected for studies comparing five immunological labeling methods using human IgG as a model antigen. Results were evaluated on the basis of the least amount of detectable antigen and background staining. The most sensitive dot-blot method was then tested for its applicability to Western blots. For both dot-blots and Western blots, the immunoalkaline phosphatase methods are more sensitive than the corresponding immunoperoxidase methods. The use of biotinylated secondary antibodies and an avidin-enzyme conjugate is recommended. Disclosure of alkaline phosphate is best achieved with naphthol AS phosphate as substrate and fast blue BB as chromogen. Peroxidase is best stained using H2O2 and diaminobenzidine (DAB). Potential endogenous enzyme activities are demonstrable by blotting methods but can be inhibited by including levamisole in the disclosure reaction medium for calf intestinal alkaline phosphatase indicators, or by incubation of blots with sodium azide and hydrogen peroxide before immunolabeling when using horseradish peroxidase indicators.

One-step automated alkaline phosphatase analysis

1966 ◽  
Vol 14 (2) ◽  
pp. 263-269 ◽  
Author(s):  
Darlene Comfort ◽  
D.J. Campbell
Keyword(s):  
Alkaline Phosphatase ◽  
One Step

scholarly journals New chromogens for alkaline phosphatase histochemistry: salmon and magenta phosphate are useful for single- and double-label immunohistochemistry.

1994 ◽  
Vol 42 (4) ◽  
pp. 551-554 ◽  
Author(s):  
C Avivi ◽  
O Rosen ◽  
R S Goldstein
Keyword(s):  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Molecular Biology ◽  
Horseradish Peroxidase ◽  
Reaction Products ◽  
Immunocytochemical Detection ◽  
Hematoxylin Staining ◽  
Double Label ◽  
Biology Research

Two new substrate chromogens for alkaline phosphatase (ALP) detection have been recently synthesized for use in molecular biology research, salmon and magenta phosphate. We show here that these two chromogens have advantageous characteristics for immunocytochemistry as well. Their relatively delicate pink- and magenta-colored products do not mask the colors produced by other staining procedures. In addition, the reaction products of these substrates are insoluble in water, ethanol, and xylene, permitting the use of regressive hematoxylin staining procedures and coverslipping in permanent resin-based media. Most importantly, when these ALP substrates are used in double-label immunocytochemistry in combination with horseradish peroxidase-diaminobenzidine (HRP-DAB) and counterstained with hematoxylin, all three colors can be easily distinguished. An application using these substrates for simultaneous immunocytochemical detection of two monoclonal antibodies of different classes, in combination with hematoxylin staining, is illustrated.

scholarly journals Reaction Rate Retardation as a Method for Serum Alkaline Phosphatase Isoenzyme Measurement

1980 ◽  
Vol 17 (4) ◽  
pp. 192-198 ◽  
Author(s):  
Pamela B Brown ◽  
K O Lewis
Keyword(s):  
Alkaline Phosphatase ◽  
Reaction Rate ◽  
Serum Alkaline Phosphatase ◽  
Enzyme Reaction ◽  
Intestinal Alkaline Phosphatase ◽  
Wide Range ◽  
Alkaline Phosphatase Isoenzymes ◽  
Sensitivity And Selectivity ◽  
Rate Retardation ◽  
Alkaline Phosphatase Isoenzyme

A method for serum alkaline phosphatase isoenzymes using an enzyme reaction rate analyser is described. The complete urea-induced degradation of enzyme activity is monitored, from which individual isoenzyme activities are obtained by calculating the constituent exponential components of the degradation curve. Activities have been measured with adequate sensitivity and selectivity for up to four isoenzyme components in normal and in pathological sera. The identity of each isoenzyme present is assigned from its characteristic degradation half-life, and by this method bone and liver alkaline phosphatase are clearly distinguished and quantitated, and a composite value for placental-intestinal alkaline phosphatase activity is obtained. The approach promises to be applicable to a wide range of isoenzymes, and in analogy with ‘reaction rate’ the term ‘reaction rate retardation’ is suggested for the procedure.

scholarly journals Development of a one-step immunoassay for triazophos using camel single-domain antibody–alkaline phosphatase fusion protein

2019 ◽  
Vol 411 (6) ◽  
pp. 1287-1295 ◽  
Author(s):  
Kai Wang ◽  
Zhiping Liu ◽  
Guochun Ding ◽  
Ji Li ◽  
Natalia Vasylieva ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Fusion Protein ◽  
Single Domain ◽  
Single Domain Antibody ◽  
Domain Antibody ◽  
One Step

Reactions in inclusion molecular complexes. A one-step regiospecific and stereospecific hydroxylation of deoxycholic acid

1975 ◽  
pp. 864 ◽  
Author(s):  
Noga Friedman ◽  
Meir Lahav ◽  
Leslie Leiserowitz ◽  
Ronit Popovitz-Biro ◽  
Chia-Pin Tang ◽  
...  
Keyword(s):  
Deoxycholic Acid ◽  
Molecular Complexes ◽  
One Step

Spark Plasma Synthesis/Sintering of Dense Ceramic, Intermetallic and Composite Materials

2006 ◽  
Vol 45 ◽  
pp. 1411-1416
Author(s):  
Antonio Mario Locci ◽  
Roberta Licheri ◽  
Roberto Orrù ◽  
A. Cincotti ◽  
Giacomo Cao
Keyword(s):  
Mechanical Load ◽  
Plasma Synthesis ◽  
Simultaneous Application ◽  
Plasma Sintering ◽  
Pulsed Dc ◽  
Spark Plasma ◽  
One Step ◽  
Powder Particles ◽  
Dc Current ◽  
Spark Plasma Synthesis

Spark Plasma Sintering (SPS) represents a very attractive technique for the obtainment of dense materials including nanostructured ones. SPS basically consists in the simultaneous application of a pulsed DC current and an uniaxial mechanical load through a powder compact. Other than providing rapid Joule heating and likely enhancing mass transport through electromigration, the imposed pulsed high current is also reported to generate a plasma within the voids surrounding the powder particles, thus facilitating the removal of oxides surface layers that may hinder the sintering process. Selected results obtained through SPS in our laboratory for the preparation of a wide variety of materials, i.e. TiC-TiB2, MgB2, and NbAl3, will be presented in this work. Specifically, all the chosen examples are related to the use of the SPS technique for obtaining the desired material by simultaneously performing synthesis and consolidation stages in one-step.

scholarly journals A comparative study of horseradish peroxidase conjugates prepared with a one-step and a two-step method.

1975 ◽  
Vol 23 (3) ◽  
pp. 200-207 ◽  
Author(s):  
D M Boorsma ◽  
G L Kalsbeek
Keyword(s):  
Horseradish Peroxidase ◽  
Lupus Erythematosus ◽  
Cross Linking ◽  
Discoid Lupus Erythematosus ◽  
Gel Chromatography ◽  
Ammonium Sulfate Precipitation ◽  
Step Method ◽  
One Step ◽  
Cryostat Sections ◽  
Unfixed Cryostat

In this study we compared horseradish peroxidase (HRP)-labeled rabbit antihuman immunoglobulin G (IgG) conjugates, prepared by a one-step and a two-step method. Glutaraldehyde was used as a cross-linking agent. Two methods were used for removing unconjugated HRP: Sephadex G-200 gel chromatography and ammonium sulfate precipitation. The conjugates were characterized immunologically, immunochemically and enzymatically. The immunohistoenzymic properties of the conjugates were tested on unfixed cryostat sections of the skin of patients with chronic discoid lupus erythematosus. The influence of the presence of unconjugated HRP and unconjugated IgG was studied. Optimal results were obtained with conjugates prepared by a two-step method. Removing unconjugated HRPimproved the immunohistoenzymic properties of the conjugates. Conjugated and unconjugated IgG could be separated by Sephadex G-200 gel chromatography.

Simultaneous Synthesis and Consolidation of Nanostructured ZrB2–ZrO2 Composite

2020 ◽  
Vol 20 (7) ◽  
pp. 4349-4352
Author(s):  
Seong-Eun Kim ◽  
Jin-Kook Yoon ◽  
In-Jin Shon
Keyword(s):  
Mechanical Properties ◽  
Combustion Synthesis ◽  
High Frequency ◽  
Combined Effects ◽  
Induced Current ◽  
Mechanical Pressure ◽  
Simultaneous Application ◽  
Average Grain Size ◽  
One Step ◽  
Simultaneous Synthesis

A dense nanostructured 2ZrB2–ZrO2 composite was synthesized by the high-frequency inductionheated combustion synthesis (HFIHCS) method within 2 min in one step from mechanically activated powders of 2B2O3 and 3Zr. Simultaneous combustion synthesis and densification were accomplished under the combined effects of the induced current and mechanical pressure. A highly dense 2ZrB2–ZrO2 composite with relative density of up to 95.5% was produced under the simultaneous application of a pressure of 80 MPa and the induced current. The average grain size and mechanical properties (hardness and fracture toughness) of the composite were investigated.

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