PARP1 Automodified Protein

PARP1 Automodified, human lyophilized

Cat. Num. AAA376019

• Applications: Western Blot, SDS-Page, ELISA
• Synonyms: PARP1 Automodified; human lyophilized; PARP1 Automodified protein

• Host: Human
• Form/Format: 5 ug PARP1 automodified, human, lyophilized.

• Applicable Applications for PARP1 Automodified protein: Western Blot (WB), SDS-PAGE standard, ELISA (EIA)
• Application Notes: Western Blot, 4 uL/well (25 ug/mL in sample buffer; 100 ng) using HRP/TMB colorimetric methods; <1 uL/well for ECL. SDS-PAGE standard, blue-stained, 30 uL per well (25 ug/mL in sample buffer; 750 ng) ELISA, 10 ng/well; To the reconstituted PARP1 automodified, prepared as above, add an equal volume of SDS-PAGE 2x sample buffer containing reducing agent (betaME or DTT). Vortex, heat at 90 degree C for 2 minutes, then spin briefly. Store at -20 or -70 degree C. Final PARP1 concentration will be 25 ug/mL.
• Reconstitution: Spin tube in a microfuge for 15 sec to sediment material to the bottom of the microfuge tube. Carefully open vial and add 100 uL dH2O. Let 0set for 5 min. Vortex gently for 1 min, avoiding air bubbles. Spin briefly in microfuge to consolidate. Upon reconstitution with 100 uL dH20, the final concentrations are as follows: 50 ug/mL PARP automodified, 50 mM Tris, pH8, 100 mM NaCl, 10 mM MgCl2, 1 mM DTT, 1 mM NAD+, 10 ug/mL activated DNA, plus lyophilization stabilizers. Store reconstituted PARP1 automodified at -70 degree C. NOTE:Reconstituted PARP1 will retain residual poly ADP-ribosylation enzymatic activity.
• Preparation and Storage: Stable for >6 months from date of shipment when stored desiccated at 4 degree C.

Testing Data

Testing Data #2

Related Product Information for PARP1 Automodified protein

Background: Poly-ADP-ribose metabolism plays a mayor role in a wide range of biological processes, such as maintenance of genomic stability, transcriptional regulation, energy metabolism and apoptosis. At least six poly-ADP-ribose polymerase (PARP) mammalian enzymes have been identified: PARP1, PARP2, PARP3, PARP4/vPARP, PARP5/Tankyrases-1 and PARP6/Tankyrases-2. Poly(ADP-ribose) polymerase-1 (PARP1) is an abundant and ubiquitous nuclear enzyme that catalyzes the NAD(+)-dependent addition of ADP-ribose polymers on a variety of nuclear proteins. PARP1 Automodified is highly purified human PARP1, expressed in a baculovirus expression system, to which NAD+ and cofactors have been added to allow PARP automodification by poly(ADP-ribose). The PARP1 is lyophilized from its native state, and upon reconstitution with water, it retains residual poly ADP-ribosylation enzymatic activity. It is useful as a standard for ELISA, SDS-PAGE, Western blotting, and in other assays.

References

D. Lamarre et al. (1988) Biochim. Biophys. Acta 950 147 Y.A. Lazebnik et al. (1994) Nature (Lond.) 371 346 P.J. Duriez et al. (1997) Biochim. Biophys. Acta 1334 65 S. Gobeil et al. (2001) Cell Death Different. 8 588 F.R. Sallmann et al. (1997) Biochem. Cell Biol. 75 451 B.C. Woodhouse and G.L. Dianov (2008) DNA Repair (Amst.) 7 1077 W.L. Kraus (2008) Curr. Opin. Cell Biol. 20 294 P.O. Hassa and M.O. Hottiger (2008) Frontiers Bioscience 13 3046
Y. Zhang et al. (2013) Nature Methods 10, 981