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P nitrophenyl thymidine 5 monophosphate

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P-nitrophenyl thymidine 5′-monophosphate is a laboratory reagent used in biochemical research. It serves as a substrate for the detection and quantification of enzymatic activity.

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Close spelling variants of this product

Sodium thymidine 5'-monophosphate p-nitrophenyl ester (nitrophenyl-pT), Thymidine 5'-monophosphate p-nitrophenyl ester Thymidine 5′ – monophosphate p-nitrophenyl ester sodium salt Thymidine

2 protocols using p nitrophenyl thymidine 5 monophosphate

1

Regulation of ENPP1 Activity in MC3T3 Cells

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MC3T3 cells were transfected with Il15ra shRNA vectors (OriGene) and differentiated for 5 days. Nucleoside triphosphate pyro-phosphohydrolase (NTPPPH) enzymatic activity correlates with ENPP1 activity and was assayed using the colorimetric substrate, p-nitrophenyl thymidine 5′-monophosphate (Sigma) as previously described[33 (link)]. Alkaline phosphatase (ALP) activity was assayed using p-nitrophenyl phosphate (pNPP) as substrate. To detect IL15RA protein levels, MC3T3 were lysed with RIPA buffer supplemented with protease inhibitors. After a pre-clearing step, lysates were incubated with a rat anti-IL15RA antibody (AbCam), followed by a precipitation step using magnetic beads. After SDS gel electrophoresis and western blotting, IL15RA was detected using a goat anti-IL15RA antibody (R&D clone af551) and a trueblot anti-goat secondary antibody (Rockland).
Loro E., Ramaswamy G., Chandra A., Tseng W.J., Mishra M.K., Shore E.M, & Khurana T.S. (2017). IL15RA is required for osteoblast function and bone mineralization. Bone, 103, 20-30.
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2

Proliferation Assay in Transfected Cells

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Transfected CH-8 cells were harvested as pellets via scraping at 24, 48 and 72 hours. The colourimetric substrate p-nitrophenyl thymidine 5′-monophosphate (Sigma, Welwyn Garden City, UK) was assessed following a published protocol24 (link).
Vijen S., Hawes C., Runions J., Russell R.G., Wordsworth B.P., Carr A.J., Pink R.C, & Zhang Y. (2020). Differences in intracellular localisation of ANKH mutants that relate to mechanisms of calcium pyrophosphate deposition disease and craniometaphyseal dysplasia. Scientific Reports, 10, 7408.
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