Product nameHuman Cleaved PARP1 ELISA Kit
Intra-assay Sample n Mean SD CV% Overall 5 6.6% Inter-assay Sample n Mean SD CV% Overall 3 8.5%
Sample typeCell culture extracts
Assay typeSandwich (quantitative)
Range7.8 µg/ml - 500 µg/ml
Sample specific recovery Sample type Average % Range Cell culture media 59 43% - 73% Fetal Bovine Serum 65 52% - 85% Bovine Serum Albumin 75 57% - 94%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Human Cleaved PARP1 SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of cleaved PARP1 protein in human cell extracts. Quantitate human cleaved PARP1 with 5.8 µg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.
PARP1 is a 113 kDa nuclear DNA-repair enzyme that transfers ADP-ribose units from NAD+ to variety of nuclear proteins including topoisomerases, histones and PARP1 itself. Via poly ADP ribosylation, PARP1 is responsible for regulation of cellular homeostasis including cellular repair, transcription and replication of DNA, cytoskeletal organization and protein degradation. In response to DNA damage, PARP1 activity is increased upon binding to DNA strand nicks and breaks. Excessive DNA damage leads to generation of large branched ADP-ribose polymers and activation of a unique cell death program.
During apoptosis, PARP1 is cleaved by activated caspase-3 between Asp214 and Gly215, resulting in the formation of an N-terminal 24 kDa fragment containing most of the DNA binding domain and a C-terminal 89 kDa fragment containing the catalytic domain. The proteolysis of PARP1 through this cleavage renders the enzyme inactive and this further facilitates apoptotic cell death. Thus the presence of 89 kDa PARP1 fragment is considered to be a very reliable biomarker of apoptosis.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Human Cleaved PARP1 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5B 1 x 6ml HeLa (Staurosporin-treated) Lyophilized Extract 2 x 250µg HeLa (untreated) Lyophilized Extract 2 x 250µg Human Cleaved PARP1 Capture Antibody (Lyophilized) 1 x 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
- Epigenetics and Nuclear Signaling
- DNA / RNA
- DNA Damage & Repair
- DNA Damage Response
- DNA Damage Recognition
FunctionInvolved 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. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. 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. Required for PARP9 and DTX3L recruitment to DNA damage sites. PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites.
Sequence similaritiesContains 1 BRCT domain.
Contains 1 PARP alpha-helical domain.
Contains 1 PARP catalytic domain.
Contains 2 PARP-type zinc fingers.
modificationsPhosphorylated by PRKDC and TXK.
Poly-ADP-ribosylated by PARP2. Poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites.
S-nitrosylated, leading to inhibit transcription regulation activity.
Cellular localizationNucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
- Information by UniProt
- ADP ribosyltransferase diphtheria toxin like 1
- ADP ribosyltransferase NAD(+)
- ADP-ribosyltransferase diphtheria toxin-like 1
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Background-subtracted data values (mean +/- SD) are graphed.
Untreated and staurosporine (ab120056) treated HeLa and Jurkat lysates were prepared in 1X Cell Extraction Buffer PTR and tested using the Cleaved PARP1 SimpleStepELISA. Raw OD 450 nm values are shown for 500 µg/mL lysate loads.
HeLa cells were treated with a dose titration of Staurosporine for 4 hours in complete media. Cells were cultured and treated in a 96-well cell culture microtiter plate. Lysates were prepared by direct in-well lysis without media removal: 2X Cell Extraction Buffer PTR was added to an equal volume of media and then resulting lysate was used directly in the Cleaved PARP1 SimpleStepTM ELISA assay. Raw values for triplicate measurements are plotted. The calculated IC50 is 0.77 µM.
20 µg of HeLa extracts that were untreated or treated for 4 hours with 1 µM Staurosporine were analyzed by western blot. The GAPDH blot is included to show the relative loads of each lysate. In the HeLa cell line, Staurosporine treatment is required to detect cleaved PARP1 protein, as observed in the SimpleStep ELISA (Figure 2).
This product has been referenced in:
- Zhu J et al. Nimotuzumab enhances the sensitivity of non-small cell lung cancer cells to tumor necrosis factor-a by inhibiting the nuclear factor-?B signaling pathway. Exp Ther Med 15:3345-3351 (2018). Read more (PubMed: 29545853) »
- de Oliveira MR et al. Pinocembrin Suppresses H2O2-Induced Mitochondrial Dysfunction by a Mechanism Dependent on the Nrf2/HO-1 Axis in SH-SY5Y Cells. Mol Neurobiol 55:989-1003 (2018). Read more (PubMed: 28084593) »