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 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 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

scholarly journals One-Step Immunoassay for Tetrabromobisphenol A Using a Camelid Single Domain Antibody–Alkaline Phosphatase Fusion Protein

2015 ◽  
Vol 87 (9) ◽  
pp. 4741-4748 ◽  
Author(s):  
Jia Wang ◽  
Zuzana Majkova ◽  
Candace R. S. Bever ◽  
Jun Yang ◽  
Shirley J. Gee ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Fusion Protein ◽  
Single Domain ◽  
Tetrabromobisphenol A ◽  
Single Domain Antibody ◽  
Domain Antibody ◽  
One Step

scholarly journals Double-enzyme conjugates, producing an intermediate color, for simultaneous and direct detection of three different intracellular immunoglobulin determinants with only two enzymes.

1986 ◽  
Vol 34 (4) ◽  
pp. 423-428 ◽  
Author(s):  
E Claassen ◽  
D M Boorsma ◽  
N Kors ◽  
N Van Rooijen
Keyword(s):  
Alkaline Phosphatase ◽  
Staining Method ◽  
Direct Detection ◽  
Direct Method ◽  
Simultaneous Detection ◽  
Single Section ◽  
Mouse Immunoglobulin ◽  
Enzyme Conjugate ◽  
One Step ◽  
Triple Staining

A new double-enzyme conjugate was synthesized by coupling alkaline phosphatase (AP) to horseradish peroxidase (HRP). After AP (blue) and subsequent HRP (red) cytochemistry, this new conjugate produced a stable intermediate-colored (violet) product. By coupling this double-enzyme conjugate to an antigen (trinitrophenyl, TNP) or an antibody (anti-mouse immunoglobulin G2a), anti-TNP or -IgG2a-producing cells could be demonstrated as violet cells in spleen sections. This led to the development of a rapid one-step incubation--two-step cytochemical procedure for simultaneous detection of three different determinants in a single tissue section. To demonstrate this novel triple staining method, we coupled three different antigens to, respectively, AP, HRP, and AP-HRP. When spleen sections of immunized animals were incubated with a mixture of these three antigen-enzyme conjugates, we could distinguish antibody-forming cells against each of these three antigens simultaneously as red (HRP), blue (AP), and violet (AP-HRP) cells. The simultaneous detection of three different classes of intracellular antibodies in a single section also proved to be possible with this method. With this study we provide a new direct method for detection of three different intracellular immunoglobulins after a one-step incubation and a two-step standard cytochemical procedure.

scholarly journals Nanobody-Alkaline Phosphatase Fusion Protein-Based Enzyme-Linked Immunosorbent Assay for One-Step Detection of Ochratoxin A in Rice

Sensors ◽  
2018 ◽  
Vol 18 (11) ◽  
pp. 4044 ◽  
Author(s):  
Zhichang Sun ◽  
Xuerou Wang ◽  
Qi Chen ◽  
Yonghuan Yun ◽  
Zongwen Tang ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Fusion Protein ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Limit Of Detection ◽  
Binding Capacity ◽  
Cross Reactivity ◽  
Solvent Tolerance ◽  
Enzyme Linked Immunosorbent Assay ◽  
One Step

Ochratoxin A (OTA) has become one a focus of public concern because of its multiple toxic effects and widespread contamination. To monitor OTA in rice, a sensitive, selective, and one-step enzyme-linked immunosorbent assay (ELISA) using a nanobody-alkaline phosphatase fusion protein (Nb28-AP) was developed. The Nb28-AP was produced by auto-induction expression and retained an intact antigen-binding capacity and enzymatic activity. It exhibited high thermal stability and organic solvent tolerance. Under the optimal conditions, the developed assay for OTA could be finished in 20 min with a half maximal inhibitory concentration of 0.57 ng mL−1 and a limit of detection of 0.059 ng mL−1, which was 1.1 times and 2.7 times lower than that of the unfused Nb28-based ELISA. The Nb28-AP exhibited a low cross-reactivity (CR) with ochratoxin B (0.92%) and ochratoxin C (6.2%), and an ignorable CR (<0.10%) with other mycotoxins. The developed Nb-AP-based one-step ELISA was validated and compared with a liquid chromatography-tandem mass spectrometry method. The results show the reliability of Nb-AP-based one-step ELISA for the detection of OTA in rice.

One-step ultrasonic synthesis of graphene quantum dots with high quantum yield and their application in sensing alkaline phosphatase

2015 ◽  
Vol 51 (5) ◽  
pp. 948-951 ◽  
Author(s):  
Yanhong Zhu ◽  
Guangfeng Wang ◽  
Hong Jiang ◽  
Ling Chen ◽  
Xiaojun Zhang
Keyword(s):  
Quantum Dots ◽  
Alkaline Phosphatase ◽  
Graphene Oxide ◽  
Quantum Yield ◽  
Graphene Quantum Dots ◽  
Label Free ◽  
High Quantum Yield ◽  
Ultrasonic Synthesis ◽  
High Quantum ◽  
One Step

With only graphene oxide and KMnO4, the luminescent graphene quantum dots (GQDs) in high quantum yield were prepared by one-step synthesis using ultrasonication, and applied in the label-free, simple and fast fluorescence assay of alkaline phosphatase (ALP).

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