Human Poly (ADP-ribose) Polymerase-1

Mfr.No.	PARP-100
Description	Protein construct: Wild-type Human Poly (ADP-ribose) Polymerase-1 purified from an insect protein expression system. MW: 120 kDa Enzyme concentration: 2 µM Enzyme assay: The kinase activity of Human Poly (ADP-ribose) Polymerase-1 is measured by using the Human Poly (ADP-ribose) Polymerase-1 Assay Kit (Catalog No. PAR100K). Storage temperature: -20 or -80°C. Do not freeze-and-thaw repeatedly. Enzyme dilution: Use the 1 x assay to dilute the enzyme just before the assay as needed. Do not store diluted enzyme solution. The Human Poly (ADP-ribose) Polymerase-1– for 100 assays (Catalog No: PARP-100) includes 17 µl of 300 x Human Poly (ADP-ribose) Polymerase-1. It is for 100 assays.

Product Manual	Datasheet
Protocols Description	Assay Protocol using the Human Poly (ADP-ribose) Polymerase-1 Assay Kit 1. Reagent preparation: 10 x DNA: dilute the 100 x DNA 10 –fold with water 10 x enzyme: dilute the 400 x Human PARP-1 40 –fold with 1 x Buffer 1 mM NAD+ : dilute the 10 mM stock 10 – fold with water 1 x fluorescence dye: dilute the 100 x fluorescence dye 100-fold with water 2. Reaction: The total volume of each reaction mixture is 50 µl including: 30 µl of H2O, 5 µl of 10 x buffer, 5 µl of 10 x DNA, 5 µl of 10 x human PARP-1, 5 µl of 1 mM NAD+ . Incubate the reaction mixture at 37°C for 60 min. 3. Detection: Add 200 µl of the 1 x fluorescence dye into the 50 µl of the reaction mixture. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.

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