Pnpp liquid substrate system for elisa

Manufactured by Merck Group

Sourced in United States

PNpp) Liquid Substrate (System for ELISA) is a laboratory product used in enzyme-linked immunosorbent assay (ELISA) procedures. It provides a liquid substrate for the detection and quantification of target analytes in samples. The product's core function is to facilitate the colorimetric detection step in ELISA protocols.

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PNPP substrate (69 citations) ELISAs (4 citations) Alkaline Phosphatase Yellow (pNPP) Liquid Substrate System for ELISA (4 citations) Liquid substrate system for ELISA (3 citations) PNPP) Liquid Substrate System (3 citations) Substrates (2 citations)

2 protocols using pnpp liquid substrate system for elisa

Alkaline Phosphatase Activity Assay

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50,000 cells were seeded in 24-well plates. When confluent media was changed to differentiation media (compare Differentiation and alkaline phosphatase staining) and the first measurement was performed. After CTB as described above, wells were PBS washed and fixated for 60 s with 4% PFA in PBS. After PBS washing 500 µl Alkaline Phosphatase Yellow (pNpp) Liquid Substrate (System for ELISA, Sigma-Aldrich, USA) was added and incubated at 37 °C for 8 min. Absorbance was measured at 405 nm.

Lutsik P., Baude A., Mancarella D., Öz S., Kühn A., Toth R., Hey J., Toprak U.H., Lim J., Nguyen V.H., Jiang C., Mayakonda A., Hartmann M., Rosemann F., Breuer K., Vonficht D., Grünschläger F., Lee S., Schuhmacher M.K., Kusevic D., Jauch A., Weichenhan D., Zustin J., Schlesner M., Haas S., Park J.H., Park Y.J., Oppermann U., Jeltsch A., Haller F., Fellenberg J., Lindroth A.M, & Plass C. (2020). Globally altered epigenetic landscape and delayed osteogenic differentiation in H3.3-G34W-mutant giant cell tumor of bone. Nature Communications, 11, 5414.

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Quantifying Alkaline Phosphatase Activity in Extracellular Vesicles

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The activity of tissue-nonspecific alkaline phosphatase (TNAP, UniProtKB Q92058) present in the MV membrane was measured using the ALP Yellow para-nitrophenyl phosphate (pNPP) Liquid Substrate System for ELISA (Sigma-Aldrich) [32 (link)]. Aliquots (10 μl) of MVs (~20 μg total protein/ml) were mixed with 200 μl of p-NPP into a 96-well plate at 37°C for 5 min, and the absorbance was recorded at 405 nm for 200 min using a microtiter plate reader. Three independent measurements were performed. TNAP activity was calculated as the number of enzyme units (U) per mg of total protein (1 U corresponds to 1 μmol of pNPP hydrolyzed per min).

Plaut J.S., Strzelecka-Kiliszek A., Bozycki L., Pikula S., Buchet R., Mebarek S., Chadli M., Bolean M., Simao A.M., Ciancaglini P., Magrini A., Rosato N., Magne D., Girard-Egrot A., Farquharson C., Esener S.C., Millan J.L, & Bottini M. (2019). Quantitative Atomic Force Microscopy Provides New Insight into Matrix Vesicle Mineralization. Archives of biochemistry and biophysics, 667, 14-21.

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