PARP1 Enzyme human Protein

PARP1 Enzyme human, lyophilized

Cat. Num. AAA376020

• Applications: Western Blot, SDS-Page, ELISA
• Purity: Purity >95% by SDS-PAGE.
• Synonyms: PARP1 Enzyme human; lyophilized; PARP1 Enzyme human protein

• Purity/Purification: Purity >95% by SDS-PAGE.
• Form/Format: 50 ug PARP1 enzyme, human, lyophilized. When reconstituted with 100 uL H2O, concentration will be 0.5 mg/mL (see below for reconstitution instructions).

• Applicable Applications for PARP1 Enzyme human protein: Western Blot (WB), SDS-PAGE standard, ELISA (EIA)
• Application Notes: Enzyme activity/inhibition assays Western Blot, 100 ng/well using HRP/TMB; <20 ng/well for ECL. SDS-PAGE standard, CB stained, 1 ug per well ELISA, 10 ng/well; To the reconstituted PARP1 enzyme, 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.; 1000 U/vial, specific activity=20000 U/mg PARP1. 1U=10 fmol ADP-ribose incorporated into 5 ug immobilized histone in 30 min at room temperature. Note: Activity measurements are approximate values.
• Reconstitution: Spin tube in a microfuge for 15 sec to sediment lyophilized material. Carefully open vial and add 100 uL dH2O. Vortex gently for 20 sec (avoid air bubbles). Let stand for 5 min. Carefully triturate the sample 10-times using a pipetman (avoid air bubbles). Spin briefly in microfuge to consolidate. Upon reconstitution with 100 uL dH20, the final concentrations are as follows: 0.5 mg/mL PARP1 enzyme, 20 mM Tris, pH8, 0.3 M NaCl, 0.1 mM EDTA, 1 mM DTT, plus lyophilization stabilizers. Store reconstituted PARP1 enzyme at -70 degree C. CAUTION: There is loss of PARP enzymatic activity upon each free/thaw cycle. It is suggested to aliquot the reconstituted enzyme into multiple tubes and freeze at -70 degree C. Alternatively, add glycerol at 1:1 vol/vol to the reconstituted PARP1, mix gently by trituration, store at -20 degree C (do not store in a frost-free freezer!) for up to 6 months.
• Preparation and Storage: Stable for 6 months from date of shipment when stored desiccated at 4 degree C, or more stable at -70 degree C.

Testing Data

Testing Data #2

Testing Data #3

Related Product Information for PARP1 Enzyme human 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.
Product Description: PARP1 Enzyme is highly purified and enzymatically active human PARP1, expressed in a baculovirus expression system. Upon addition of NAD+ and cofactors the PARP1 is automodified by the addition of ADP-ribose monomers to form poly(ADP-ribose) polymer. The PARP1 is supplied lyophilized, and upon reconstitution with water, it retains its native poly ADP-ribosylation enzymatic activity. It is useful for high-throughput enzymatic assays, visualization of the automodification reaction by SDS-PAGE and Western blotting, ELISA, standard for SDS-PAGE and WB, and in other assays.

References

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