Invitrogen

PARP1 (cleaved Asp214, Asp215) Monoclonal Antibody (194C1439)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

View all (14) PARP1 (cleaved Asp214) antibodies

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PARP1 (cleaved Asp214, Asp215) Antibody in Western Blot (WB) — CRISPR-Cas9 mediated genome editing of PARP1 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR978664_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of PARP1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), Treated HeLa Wild Type (Lane 2), HeLa Cas9 (Lane 3), Treated HeLa Cas9 (Lane 4), HeLa Cas9 cells transduced with PARP1 Lentiviral sgRNA (Lane 5) and Treated HeLa Cas9 cells transduced with PARP1 Lentiviral sgRNA (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-PARP1 (cleaved Asp214, Asp215) Monoclonal Antibody (194C1439) (Product # MA1-41020) using 1:1,000 dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:4,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Even though NGS analysis determine the clone as partial KO, there was complete loss of signal in sgRNA transduced cells confirming that the antibody is specific to PARP1. An uncharacterized band was observed at 49 kDa in HeLa Wild Type and Cas9 untreated samples. Treatment

PARP1 (cleaved Asp214, Asp215) Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A reduced signal was observed for target protein in HeLa Cas9 cell line transduced with PARP1 Lentiviral sgRNA compared to control cell line using Anti-PARP1 (cleaved Asp214, Asp215) Monoclonal Antibody (194C1439) (Product # MA1-41020). {KO}

Product Details

MA1-41020

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	-
Flow Cytometry (Flow)	1-3 µg/test	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Mouse / IgG2b, kappa
Class	Monoclonal
Type	Antibody
Clone	194C1439
Immunogen	Synthetic peptide corresponding to residues near the 214/215-cleavage site of human PARP. View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	1.0 mg/mL
Purification	Protein G
Storage buffer	PBS
Contains	0.05% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_1077468

Product Specific Information

Suggested positive control: camptothecin treated HL-60 (12 hr).

Target Information

Poly ADP-Ribose Polymerase (PARP) uses nicotinamide adenine dinucleotide (oxidized form) NAD as a substrate to catalyse the transfer of ADP-ribose to a variety of nuclear protein acceptors. Proteolysis of PARP to its stable 85kDa fragment is an early marker of programmed cell death (apoptosis) and is mediated by the caspase CPP32 protein. Cleavage occurs between Adp216 and Gly217, a site in PARP conserved across species.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase); ADP-ribosyltransferase diphtheria toxin-like 1; ADP-ribosyltransferase NAD(+); ADPRT 1; ARTD1; DNA ADP-ribosyltransferase PARP1; NAD(+) ADP-ribosyltransferase 1; PARP-1; poly (ADP-ribose) polymerase 1; poly (ADP-ribose) polymerase family, member 1; Poly [ADP-ribose] polymerase 1; poly(ADP-ribose) polymerase; poly(ADP-ribose) synthetase; poly(ADP-ribosyl)transferase; Poly[ADP-ribose] synthase 1; Protein poly-ADP-ribosyltransferase PARP1

Gene Aliases: ADPRT; ADPRT 1; ADPRT1; ARTD1; pADPRT-1; PARP; PARP-1; PARP1; PPOL

UniProt ID: (Human) P09874

Entrez Gene ID: (Human) 142

Molecular Function: DNA metabolism protein