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Product name
Cleaved PARP Human ELISA Kit
Precision
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| 1 | 8 | 4.9% |
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| 1 | 4 | 10.9% |
Tests
1 x 96 well plate
Sample type
Cell culture extracts, Tissue Extracts
Assay type
Sandwich
Sensitivity
15 µg/ml
Range
8 µg/ml - 250 µg/ml
Recovery
95%
| Sample type | Average % | Range % |
|---|---|---|
| Cell culture supernatant | 93 | 82 - 109 |
| Serum | 106 | 100 - 110 |
Cross reactivity
Reacts with
Human
Notes
PARP-1 is a 113 kDa nuclear DNA-repair enzyme that transfers ADP-ribose units from NAD+ to variety of nuclear proteins including topoisomerases, histones and PARP-1 itself. Via poly ADP ribosylation, PARP-1 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, PARP-1 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, PARP-1 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 PARP-1 through this cleavage renders the enzyme inactive and this further facilitates apoptotic cell death. Thus the presence of 89 kDa PARP-1 fragment is considered to be a very reliable biomarker of apoptosis.
Storage: Store all components at 4°C. This kit is stable for at least 6 months from receipt. After reconstitution the standard should be stored at -80°C. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
In response to DNA damage, PARP-1 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, PARP-1 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 PARP-1 through this cleavage renders the enzyme inactive and this further facilitates apoptotic cell death. Thus the presence of 89 kDa PARP-1 fragment is considered to be a very reliable biomarker of apoptosis.
Storage: Store all components at 4°C. This kit is stable for at least 6 months from receipt. After reconstitution the standard should be stored at -80°C. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
Tested applications
Sandwich ELISAmore details
Properties
Storage instructions
Please see Notes section
| Components | Identifier | 1 x 96 tests |
|---|---|---|
| 10X Blocking Buffer | P/N 8209802 | 1 x 6ml |
| ab75607 - 10X Detector Antibody | P/N 8209536 | 1 x 700µl |
| 10X HRP Label | P/N 8206026 | 1 x 1ml |
| 20X Buffer | P/N 8209534 | 1 x 20ml |
| 2X Extraction Buffer | P/N 8209533 | 1 x 15ml |
| PARP Microplate | P/N 8209535 | 1 unit |
| TMB Development Solution | P/N 8206045 | 1 x 6ml |
Research areas
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> DNA Damage Response >> DNA Damage Recognition
Epigenetics and Nuclear Signaling >> DNA / RNA >> DNA Damage & Repair >> Base Excision Repair
Epigenetics and Nuclear Signaling >> Chromatin Modifying Enzymes >> ADP-ribosylation
Function
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. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150.
Sequence similarities
Contains 1 BRCT domain.
Contains 1 PARP alpha-helical domain.
Contains 1 PARP catalytic domain.
Contains 2 PARP-type zinc fingers.
Post-translational
modifications
Phosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
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 localization
Nucleus.
Target information above from: UniProt accessionP09874
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- ADPRT 1
- NAD(+) ADP-ribosyltransferase 1
- PARP-1
see all
Database links
- Entrez Gene: 142 Human
- SwissProt: P09874 Human
- Unigene: 177766 Human
Applications
Show applications key
Our Abpromise guarantee covers the use of ab119690 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Sandwich ELISA
sELISAsELISA
Cleaved PARP Human ELISA Kit images:
ELISA - Cleaved PARP Human ELISA Kit (ab119690)
Figure 1. Example standard curve. A dilution series of pellet extracts in Incubation Buffer in the working range of the assay. The lysates were prepared from a pellet of HeLa cells treated with Staurosporine.
ELISA - Cleaved PARP Human ELISA Kit (ab119690)
Figure 2. Example experimental analysis of pellet extracts prepared from HeLa (A) or Jurkat (B) cells treated with vehicle or Staurosporine. Cell extracts at varying concentrations within the working range of the assay were analyzed. Absolute signals (without zero standard subtraction) are shown. Dashed lines represent signals of zero standard (Incubation Buffer only).
ELISA - Cleaved PARP Human ELISA Kit (ab119690)
Figure 3. Example of IC50 determination. Lysates were prepared by direct in-well lysis without media removal from Jurkat and HeLa cells grown and treated with variable doses of Staurosporine in a 96-well plate. Absolute signals of Jurkat lysates (corresponding to 10X105 cells/mL) and HeLa lysates (corresponding to 2.5x105 cells/mL) are shown, respectively, in A and B.
ELISA - Cleaved PARP Human ELISA Kit (ab119690)
Figure 4. Demonstration of assay specificity by inducing PARP cleavage with Staurosporine. (A) Cleaved PARP ELISA using vehicle or Staurosporine treated HeLa extracts (250 ug/mL) or Jurkat extracts (125 ug/mL). (B) Western blot using the Cleaved PARP ELISA capture antibody (ab110315). (C) Western blot using the Cleaved PARP ELISA detector antibody (ab75607). The same extract preparations of vehicle-treated (lane1) or Staurosporine-treated (lane 2) HeLa cells, and of vehicle-treated (lane 3) or Staurosporine-treated (lane 4) Jurkat cells were used in both ELISA and Western blot analyses. The data demonstrate that the ELISA kit measures specifically the 89 kDa fragment of PARP-1. Note of complete cleavage of PARP in the Staurosporine-treated HeLa and Jurkat samples (in C).
Protocols
References for Cleaved PARP Human ELISA Kit (ab119690)
ab119690 has not yet been referenced specifically in any publications.
Publishing research using ab119690? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"