PARP1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Poly ADP Ribose Polymerase (PARP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Poly ADP Ribose Polymerase (PARP) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

ADP-ribosyltransferase (NAD+; poly (ADP-ribose) polymerase) ; ADP-ribosyltransferase diphtheria toxin-like 1; ADP-ribosyltransferase NAD (+) ; ADPRT; ADPRT 1; ADPRT1; ARTD1; DNA ADP-ribosyltransferase PARP1; NAD (+) ADP-ribosyltransferase 1; pADPRT-1; PARP; PARP-1; poly (ADP-ribose) polymerase family, member 1; poly [ADP-ribose] polymerase 1; poly (ADP-ribose) synthetase; poly (ADP-ribosyl) transferase; poly[ADP-ribose] synthase 1; PPOL; protein poly-ADP-ribosyltransferase PARP1.

Involved in the base excision repair (BER) pathway, by catalyzing the poly (ADP-ribosyl) ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks (PubMed:17177976, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:23230272) . Mediates the poly (ADP-ribosyl) ation of APLF and CHFR (PubMed:17396150) . Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150. With EEF1A1 and TXK, forms a complex that acts as a T-helper 1 (Th1) cell-specific transcription factor and binds the promoter of IFN-gamma to directly regulate its transcription, and is thus involved importantly in Th1 cytokine production (PubMed:17177976) . Required for PARP9 and DTX3L recruitment to DNA damage sites (PubMed:23230272) . PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites (PubMed:23230272) . Mediates serine ADP-ribosylation of target proteins following interaction with HPF1; HPF1 conferring serine specificity (PubMed:28190768) . Mediates the poly (ADP-ribosyl) ation of histones in a HPF1-dependent manner (PubMed:27067600) . Involved in the synthesis of ATP in the nucleus, together with NMNAT1, PARG and NUDT5 (PubMed:27257257) . Nuclear ATP generation is required for extensive chromatin remodeling events that are energy-consuming (PubMed:27257257) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Poly ADP Ribose Polymerase (PARP) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Poly ADP Ribose Polymerase (PARP) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Poly ADP Ribose Polymerase (PARP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Poly ADP Ribose Polymerase (PARP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Component Amount PARP1 Microplate 96 Wells (12 x 8 Well strips) PARP1 Lyophilized Standard 2 100X PARP1 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Component Amount Component Amount PARP1 Microplate 96 Wells (12 x 8 Well strips) PARP1 Microplate 96 Wells (12 x 8 Well strips) PARP1 Lyophilized Standard 2 PARP1 Lyophilized Standard 2 100X PARP1 HRP-Detector Anitbody 1 x 120 uL 100X PARP1 HRP-Detector Anitbody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 2 mL TMB Substrate 1 x 2 mL Stop Solution 1 x 20 mL Stop Solution 1 x 20 mL