Protocol for evaluation and validation of TLR8 antagonists in HEK-Blue cells via secreted embryonic alkaline phosphatase assay

As an important DNA 3′-phosphatase, alkaline phosphatase can repair damaged DNA caused by replication and recombination. It is essential to measure the level of alkaline phosphatase to indicate some potential diseases, such as cancer, related to alkaline phosphatase. Here, we designed a simple and fast method to detect alkaline phosphatase quantitively. When alkaline phosphatase is present, the resulting poly T-DNA with a 3′-hydroxyl end was cleaved by exonuclease I, prohibiting the formation of fluorescent copper nanoparticles. However, the fluorescent copper nanoparticles can be monitored with the absence of alkaline phosphatase. Hence, we can detect alkaline phosphatase with this turn-off strategy. The proposed method is able to quantify the concentration of alkaline phosphatase with the LOD of 0.0098 U/L. Furthermore, we utilized this method to measure the effects of inhibitor Na3VO4 on alkaline phosphatase. In addition, it was successfully applied to quantify the level of alkaline phosphatase in human serum. The proposed strategy is sensitive, selective, cost effective, and timesaving, having a great potential to detect alkaline phosphatase quantitatively in clinical diagnosis.

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Shifting nutrient-mediated interactions between algae and bacteria in a microcosm: Evidence from alkaline phosphatase assay

Microbiological Research ◽

10.1016/j.micres.2011.10.005 ◽

2012 ◽

Vol 167 (5) ◽

pp. 292-298 ◽

Cited By ~ 36

Author(s):

Huali Liu ◽

Yiyong Zhou ◽

Wenjuan Xiao ◽

Lei Ji ◽

Xiuyun Cao ◽

...

Keyword(s):

Alkaline Phosphatase ◽

Alkaline Phosphatase Assay ◽

Algae And Bacteria ◽

Phosphatase Assay

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An ultrasensitive and simple method for alkaline phosphatase assay and targeted natural compound screening in vitro

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-018-1174-8 ◽

2018 ◽

Vol 410 (21) ◽

pp. 5219-5228 ◽

Cited By ~ 4

Author(s):

Hongyan Zhou ◽

Chunyi Tong ◽

Bin Li ◽

Bin Liu ◽

Caiyun Peng ◽

...

Keyword(s):

Alkaline Phosphatase ◽

Natural Compound ◽

Simple Method ◽

Alkaline Phosphatase Assay ◽

Compound Screening ◽

Phosphatase Assay ◽

Screening In Vitro

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Alkaline Phosphatase Assay Based on the Chromogenic Interaction of Diethanolamine with 4-Aminophenol

Analytical Chemistry ◽

10.1021/acs.analchem.8b01371 ◽

2018 ◽

Vol 90 (10) ◽

pp. 6339-6345 ◽

Cited By ~ 27

Author(s):

Jian Sun ◽

Jiahui Zhao ◽

Xingfu Bao ◽

Qifeng Wang ◽

Xiurong Yang

Keyword(s):

Alkaline Phosphatase ◽

Alkaline Phosphatase Assay ◽

Phosphatase Assay

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Temperature dependence of the absorbance of alkaline solutions of 4-nitrophenyl phosphate--a potential source of error in the measurement of alkaline phosphatase activity.

Clinical Chemistry ◽

10.1093/clinchem/23.9.1541 ◽

1977 ◽

Vol 23 (9) ◽

pp. 1541-1547 ◽

Cited By ~ 4

Author(s):

C A Burtis ◽

L E Seibert ◽

M A Baird ◽

E J Sampson

Keyword(s):

Alkaline Phosphatase ◽

Thermal Response ◽

Alkaline Solutions ◽

Reaction Conditions ◽

Potential Source ◽

Alkaline Phosphatase Assay ◽

Nitrophenyl Phosphate ◽

Phosphatase Assay ◽

And Control ◽

Source Of Error

Abstract The absorbance of an alkaline solution of 4-nitrophenyl phosphate is a function of temperature. Quantitative evaluation of this phenomenon indicates that it (a) depends on the concentration of the compound and is independent of source, buffer concentration, and pH above 9.0; (b) is reversible; (c) is not a result of alkaline hydrolysis or 4-nitrophenol contamination; and (d) correlates with a temperature-induced shift of its absorbance spectrum. The phenomenon may represent a potential analytical problem in methods for alkaline phosphatase in which this compound is the substrate. If thermal equilibrium is not reached and maintained during an alkaline phosphatase assay, the thermochromic response will be included in the measured rate. The magnitude of this error depends on the thermal response and control characteristics of each particular instrument and the reaction conditions under which such an analysis is performed.

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An Automated Alkaline Phosphatase Assay with Phenolphthalein Monophosphate as Substrate

Clinical Chemistry ◽

10.1093/clinchem/13.4.281 ◽

1967 ◽

Vol 13 (4) ◽

pp. 281-289 ◽

Cited By ~ 9

Author(s):

Kirsten Hviid

Keyword(s):

Alkaline Phosphatase ◽

Normal Values ◽

Serum Alkaline Phosphatase ◽

Colorimetric Determination ◽

Alkaline Phosphatase Assay ◽

Phosphatase Assay ◽

Hydrolysis Of

Abstract The manual procedure of Babson et al. (1) for the determination of serum alkaline phosphatase has been automated. The assay is based on the colorimetric determination of phenolphthalein formed on hydrolysis of phenolphthalein monophosphate. The procedure utilizes 0.16 ml. of serum without dialysis. Blanks are required only for turbid sera. Results are compared with those obtained by the manual procedure, and data relating to sample interaction, precision, blank values, and normal values are presented.

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Collaborative Study of a New Alkaline Phosphatase Assay System for Milk

Journal of AOAC INTERNATIONAL ◽

10.1093/jaoac/51.4.802 ◽

1968 ◽

Vol 51 (4) ◽

pp. 802-807 ◽

Cited By ~ 1

Author(s):

D H Kleyn ◽

S H C Lin

Keyword(s):

Alkaline Phosphatase ◽

Statistical Study ◽

Collaborative Study ◽

Skim Milk ◽

New Method ◽

Chi Square ◽

Alkaline Phosphatase Assay ◽

Whole Milk ◽

Phosphatase Assay ◽

Tests Of Independence

Abstract A new method in which phenolphthalein monophosphate is used as substrate has been developed for determining residual alkaline phosphatase: Upon hydrolysis, phenolphthalein is released and yields a red solution that is compared visually with a standard prepared from the same milk. Seven collaborators made 168 observations that were used in a statistical study of the results. Chi-square calculations and tests of independence indicated that the described method yields results on skim milk, whole milk, and light cream that are as precise and either as accurate or more accurate than those obtained with the Scharer I method. Based on these results, it is recommended that the method be adopted as official, first action.

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