pNPP substrate buffer is supplied as a ready-to-use diluent for PNPP, Disodium Salt (sc-3720) or p-Nitrophenyl Phosphate 50X solution (sc-358718). PNPP is the substrate of choice for use with alkaline phosphatase in Enzyme Linked Immunosorbant Assay (ELISA) procedures.
NOTE: Diluent buffer only, does not contain p-Nitrophenyl Phosphate (pNPP).
1. Voller, A., et al. 1976. Bull. World Health Organ. 53: 55-65. PMID: 1085667 2. Engvall, E. 1980. Meth. Enzymol. 70: 419-439. PMID: 6999296 3. Voller, A., et al. 1986. Manual of Clinical Laboratory Immunology, 3rd Edition. In Rose, N., et al. eds., Washington, D.C. American Society for Microbiology, 106-107.
• To prepare PNPP substrate solution, dissolve 10 mg of room-temperature PNPP, Disodium Salt (sc-3720) into 10 ml of pNPP Substrate Buffer (sc-296099). Mix well and protect from light. Diluted product may be used for up to 10 hours following mixing. Discard if solution becomes yellow or turbid.
• Following incubation of antibody samples and the addition of alkaline phosphatase labeled probes, wash plates thoroughly avoiding the use of phosphate buffers. Add 100 µl of 1X PNPP substrate solution to each well and incubate 15-60 minutes. Incubation times will vary depending on the enzyme present.
• For endpoint analysis, the reaction may be stopped with 3N NaOH and read at 405 nm.
Store at 4° C
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
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pNPP Substrate Buffer
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Various publications cite the use of pNPPVarious publications cite the use of pNPP Substrate Buffer with alkaline phosphatase in ELISA procedures. pNPP substrate buffer is supplied as a ready-to-use diluent for PNPP, Disodium Salt (sc-3720) or p-Nitrophenyl Phosphate 50X solution (sc-358718). -SCBT Publication Review
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