General description
Research area: Cell Signaling
Human PARP1 (GenBank Accession No. NM_001618), full length with N-terminal GST tag, MW = 140 kDa, expressed in a Baculovirus infected Sf9 cell expression system. Poly [ADP-ribose] polymerase 1 (PARP1) belongs to the DNA-dependent nuclear enzyme superfamily. Its structure includes an N-terminal with three zinc finger DNA-binding domains, an auto-modification domain with BRCT motif and a WGR domain having conserved tryptophan, glycine, and arginine residues, and a C-terminal catalytic domain with PARP signature sequence.
Application
PARP1 Active human has been used in GST-PARP1 pulldown, to study the alterations in protein-protein interactions of sex determining region Y-box 2 (SOX2) upon O-GlcNAcylation. It has also been used in in vitro PARylation assay to investigate the effect of NR1D1 on DNA repair after damage induced by doxorubicin in breast cancer cells. It is useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Biochem/physiol Actions
PARPs comprise a set of enzymes that govern cellular processes such as DNA damage response, cell metabolism, chromatin remodeling, and transcriptional regulation. PARP1 detects and mends DNA breaks via base excision repair when DNA damage occurs. In the absence of PARP1 activity, damaged DNA accumulates, leading to impaired DNA replication.
Physical form
Formulated in 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 50% glycerol and 3 mM DTT.
Preparation Note
Thaw on ice. Upon first thaw, briefly spin tube containing enzyme to recover full content of the tube. Aliquot enzyme into single use aliquots. Store remaining undiluted enzyme in aliquots at -70°C. Note: Enzyme is very sensitive to freeze/thaw cycles.
Other Notes
One unit of PARP incorporates 100 pmoles of poly(ADP) in 1 minute (room temperature) from NAD into acid-insoluble form.
| biological source | human |
| recombinant | expressed in baculovirus infected insect cells |
| assay | ≥80% (SDS-PAGE) |
| form | aqueous solution |
| mol wt | 140 kDa |
| packaging | pkg of 10 and 20 μg |
| manufacturer/tradename | Sigma-Aldrich |
| storage condition | avoid repeated freeze/thaw cycles |
| concentration | >0.02 mg/mL |
| technique(s) | inhibition assay: suitable |
| NCBI accession no. | NM_001618 |
| UniProt accession no. | P09874 |
| application(s) | life science and biopharma |
| shipped in | dry ice |
| storage temp. | −70°C |
| Gene Information | human ... PARP1(142) |
