PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), eFluor™ 450, eBioscience™

Product Specific Information

Description: This HLNC4 monoclonal antibody reacts with human poly (ADP-ribose) polymerase (PARP1). This ubiquitous 116 kDa nuclear enzyme is involved in DNA repair. During apoptosis, active caspases -3, -6 and -7 cleave PARP1 after Asp214, thereby inactivating PARP1 and generating two apoptotic fragments sized 85 kDa and 25 kDa.

The HLNC4 antibody specifically recognizes the 85 kDa PARP1 fragment produced after cleavage and does not recognize the full-length 116 kDa protein. The following peptide was used as the immunogen: NH2-GVDEVAKKKSKKEKDC-COOH.

Applications Reported: This HLNC4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This HLNC4 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of Jurkat cells using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to BestProtocols®: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome.

Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser.

Filtration: 0.2 µm post-manufacturing filtered.

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.