A fast and sensitive colorimetric assay for IL-6 in hepatoma cells based on the production of a secreted form of alkaline phosphatase (SEAP)

1994 ◽  
Vol 170 (1) ◽  
pp. 47-56 ◽  
Author(s):  
Bernard Gregory ◽  
Rocco Savino ◽  
Gennaro Ciliberto
1998 ◽  
Vol 46 (6) ◽  
pp. 737-743 ◽  
Author(s):  
Heiner Müller ◽  
Guoli Dai ◽  
Michael J. Soares

The rat placenta expresses a family of genes related to prolactin (PRL). Target tissues and physiological roles for many members of the PRL family have yet to be determined. In this investigation we evaluated the use of an alkaline phosphatase (AP) tag for monitoring the behavior of a prototypical member of the PRL family, placental lactogen-I (PL-I). A probe was generated consisting of a fusion protein of human placental AP and rat PL-I (AP-PL-I). The AP-PL-I construct was stably expressed in 293 human fetal kidney cells, as was the unmodified AP vector that served as a control. AP activity was monitored with a colorimetric assay in conditioned medium from transfected cells. Immunoreactivity and PRL-like biological activities of the AP-PL-I fusion protein were demonstrated by immunoblotting and the Nb2 lymphoma cell proliferation assay, respectively. AP-PL-I specifically bound to tissue sections known to express the PRL receptor, including the ovary, liver, and choroid plexus. Binding of AP-PL-I to tissues was specific and could be competed with ovine PRL. The results indicate that AP is an effective tag for monitoring the behavior of PL-I and suggest that this labeling system may also be useful for monitoring the actions of other members of the PRL family.


2008 ◽  
Vol 71 (9) ◽  
pp. 1884-1888 ◽  
Author(s):  
V. KLOTZ ◽  
ART HILL ◽  
K. WARRINER ◽  
M. GRIFFITHS ◽  
J. ODUMERU

Raw milk is a well-established vehicle for the carriage of human pathogens, and many regulatory bodies have consequently mandated compulsory pasteurization as a food safety intervention. The residual activity of alkaline phosphatase (ALP) has historically been used to verify the adequacy of pasteurization of cow's milk. However, there is uncertainty on how the current ALP standards and methods of analysis can be applied to sheep's and goat's milk, which naturally contain different levels of the enzyme than that found in cow's milk. The official ALP methods applied in Canada (colorimetric assay; MFO-3) and in the United States (Fluorophos) were assessed for their ability to detect enzyme activity in raw and pasteurized milk derived from cows, sheep, and goats. The detection limit and the limit of quantitation were 0.8 and 2.02 μg/ml phenol, respectively, for the MFO-3 method and 43 and 85 mU/liter, respectively, for the Fluorophos method. The average ALP levels in raw goat's, cow's, and sheep's milk were 165, 1,562, and 3,512 μg/ml phenol, respectively. Raw milk detection limits, which correspond to raw milk phosphatase levels, were 0.051, 0.485, and 0.023% in cow's, goat's, and sheep's milk, respectively, for the MFO-3 method and 0.007, 0.070, and 0.004%, respectively, for the Fluorophos method. Although both methods can be used for ALP determination in cow's, goat's, and sheep's milk, the Fluorophos assay was superior to the colorimetric MFO-3 method based on sensitivity and time required to complete the analysis.


2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Tetiana Haniastuti ◽  
Heni Susilowati ◽  
Margareta Rinastiti

The bone of yellowfin tuna (Thunnus albacares) contains high calcium and phosphor and can be synthesized into hydroxyapatite (HA). Due to its mineral content and similarity in chemical composition with human hard tissue, HA may have potency as a pulp capping material. The aim of this in vitro study was to evaluate the viability and alkaline phosphatase (ALP) activity of dental pulp cells after exposure to HA synthesized from yellowfin tuna bone (THA). Pulp cells were isolated from human-impacted third molar. To evaluate the viability of the pulp cells, the cells were cultured and exposed to various concentrations (6.25 to 200 μg/ml) of THA for 24, 48, and 72 hours. For ALP activity assay, pulp cells were cultured with odontoblastic differentiation media and exposed to THA for 7, 11, and 15 days. ALP activity was then determined using an ALP colorimetric assay kit. Results showed that the viability of the cells was more than 91% after exposure to various concentrations of THA and the cells demonstrated normal cell morphology in all observation periods. The ALP activity test revealed that groups exposed to THA for 7, 11, and 15 days showed higher ALP activity than the control groups ( p < 0.05 ). It is concluded that THA had no cytotoxic effect on pulp cells; furthermore, it enhanced proliferation as well as ALP activity of the pulp cells.


1998 ◽  
Vol 13 (6) ◽  
pp. 643-650 ◽  
Author(s):  
KHALEQUE NEWAZ KHAN ◽  
TAKUYA TSUTSUMI ◽  
KEISUKE NAKATA ◽  
KAZUHIKO NAKAO ◽  
YUJI KATO ◽  
...  

2013 ◽  
Vol 43 ◽  
pp. 366-371 ◽  
Author(s):  
Chun Mei Li ◽  
Shu Jun Zhen ◽  
Jian Wang ◽  
Yuan Fang Li ◽  
Cheng Zhi Huang

2019 ◽  
Vol 11 (42) ◽  
pp. 5472-5477 ◽  
Author(s):  
Linjie Wang ◽  
Kun Ye ◽  
Jianming Pan ◽  
Hongwei Song ◽  
Xin Li ◽  
...  

A catalytic reaction-based colorimetric assay of alkaline phosphatase activity is developed based on oxidase-like MnO2 microspheres.


2019 ◽  
Vol 43 (11) ◽  
pp. 4525-4530 ◽  
Author(s):  
Xuemei Hu ◽  
Chaoqun Sun ◽  
Ying Shi ◽  
Yijuan Long ◽  
Huzhi Zheng

A simple and inexpensive colorimetric assay for alkaline phosphatase (ALP) and α-fetoprotein (AFP) has been established by the hydrolysis of fluorescein diphosphate (FDP).


Talanta ◽  
2017 ◽  
Vol 163 ◽  
pp. 146-152 ◽  
Author(s):  
Qiong Hu ◽  
Minhui He ◽  
Yaqi Mei ◽  
Wenjie Feng ◽  
Su Jing ◽  
...  

2020 ◽  
Vol 12 (2) ◽  
pp. 59-63
Author(s):  
Jaber Yaghini ◽  
Samaneh Khashei ◽  
Zohreh Afshari ◽  
Ahmad Mogharehabed

Background. Evaluation of salivary biomarkers is a non-invasive, convenient, and economical method for diagnosing many diseases. Evidence shows that salivary biomarkers and periodontal disease might be correlated. This study was conducted to evaluate phase I periodontal therapy’s effect on salivary concentrations of calcium, phosphorous, and alkaline phosphatase (ALP). Methods. In this descriptive, analytical study, 16 patients were selected from those referred to the Department of Oral Medicine, Faculty of Dentistry, Isfahan University of Medical Sciences, using convenience sampling. Salivary samples were collected using the drooling method. The salivary concentrations of calcium, phosphorous, and ALP were measured immediately after saliva collection, before the first phase of periodontal therapy and one month later, using a colorimetric assay. The data were analyzed with SPSS using paired t-test. P<0.05 was considered statistically significant. Results. The salivary concentrations of calcium, phosphorous, and ALP were 6.68, 20.57, and 48.31 mg/dL, respectively, before and 7.15, 22.51, and 40.37 mg/dL, respectively, after phase I periodontal therapy. There were no significant differences between the salivary levels of calcium, phosphorous, and ALP before and after phase I periodontal therapy (P>0.05). Conclusion. This study revealed that the salivary concentrations of calcium, phosphorous, and ALP remained relatively unchanged after phase I periodontal therapy.


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