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Anti-Cleaved PARP1 antibody [E51] ab32064 is a rabbit monoclonal antibody that is used in Cleaved PARP1 western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone E51 has been tried and trusted by researchers since 2006 and is cited in >500 publications
- Specificity confirmed with PARP1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available


Images

Western blot - Anti-Cleaved PARP1 antibody [E51] (AB32064), expandable thumbnail
  • Western blot - Anti-Cleaved PARP1 antibody [E51] (AB32064), expandable thumbnail
  • Western blot - Anti-Cleaved PARP1 antibody [E51] (AB32064), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (AB32064), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (AB32064), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIHC-P
Human
Tested
Not recommended
Tested
Mouse
Tested
Not recommended
Expected
Rat
Tested
Not recommended
Tested
Chinese hamster
Predicted
Not recommended
Predicted

Tested
Tested

Species

Mouse

Dilution info

1/1000 - 1/10000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Predicted
Predicted

Species

Chinese hamster

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human, Chinese hamster

Dilution info

-

Notes

-

Tested
Tested

Species

Rat

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Chinese hamster

Dilution info

-

Notes

-

Associated Products

Select an associated product type

7 products for Alternative Product

Target data

Function

Poly-ADP-ribosyltransferase that mediates poly-ADP-ribosylation of proteins and plays a key role in DNA repair (PubMed:17177976, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:23230272, PubMed:25043379, PubMed:33186521, PubMed:32028527, PubMed:26344098). Mediates glutamate, aspartate, serine or tyrosine ADP-ribosylation of proteins: the ADP-D-ribosyl group of NAD(+) is transferred to the acceptor carboxyl group of target residues and further ADP-ribosyl groups are transferred to the 2'-position of the terminal adenosine moiety, building up a polymer with an average chain length of 20-30 units (PubMed:7852410, PubMed:9315851, PubMed:19764761, PubMed:25043379, PubMed:28190768, PubMed:29954836). Serine ADP-ribosylation of proteins constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage (PubMed:33186521). Mainly mediates glutamate and aspartate ADP-ribosylation of target proteins in absence of HPF1 (PubMed:19764761, PubMed:25043379). Following interaction with HPF1, catalyzes serine ADP-ribosylation of target proteins; HPF1 conferring serine specificity by completing the PARP1 active site (PubMed:28190768, PubMed:29954836, PubMed:33186521, PubMed:32028527). Also catalyzes tyrosine ADP-ribosylation of target proteins following interaction with HPF1 (PubMed:30257210, PubMed:29954836). PARP1 initiates the repair of DNA breaks: recognizes and binds DNA breaks within chromatin and recruits HPF1, licensing serine ADP-ribosylation of target proteins, such as histones, thereby promoting decompaction of chromatin and the recruitment of repair factors leading to the reparation of DNA strand breaks (PubMed:17177976, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:23230272, PubMed:27067600). In addition to base excision repair (BER) pathway, also involved in double-strand breaks (DSBs) repair: together with TIMELESS, accumulates at DNA damage sites and promotes homologous recombination repair by mediating poly-ADP-ribosylation (PubMed:26344098, PubMed:30356214). Mediates the poly(ADP-ribosyl)ation of a number of proteins, including itself, APLF and CHFR (PubMed:17396150, PubMed:19764761). In addition to proteins, also able to ADP-ribosylate DNA: catalyzes ADP-ribosylation of DNA strand break termini containing terminal phosphates and a 2'-OH group in single- and double-stranded DNA, respectively (PubMed:27471034). 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). Acts as a regulator of transcription: positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150 (PubMed:19344625). Plays a role in the positive regulation of IFNG transcription in T-helper 1 cells as part of an IFNG promoter-binding complex with TXK and EEF1A1 (PubMed:17177976). 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).

Alternative names

Poly [ADP-ribose] polymerase 1, PARP-1, ADP-ribosyltransferase diphtheria toxin-like 1, DNA ADP-ribosyltransferase PARP1, NAD(+) ADP-ribosyltransferase 1, Poly[ADP-ribose] synthase 1, Protein poly-ADP-ribosyltransferase PARP1, ARTD1, ADPRT 1, PPOL, ADPRT, PARP1

Recommended products

Anti-Cleaved PARP1 antibody [E51] ab32064 is a rabbit monoclonal antibody that is used in Cleaved PARP1 western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone E51 has been tried and trusted by researchers since 2006 and is cited in >500 publications
- Specificity confirmed with PARP1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

E51

Purification technique

Affinity purification Protein A

Specificity

This antibody is specific for the p25 cleaved form of human PARP1.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Cleaved PARP1 also known as cPARP is a fragment of the PARP1 protein an important DNA repair enzyme. The full PARP1 protein has a molecular weight of approximately 116 kDa but after cleavage during apoptosis the cleaved PARP1 fragments typically have a molecular weight of around 89 kDa and 24 kDa. PARP1 is expressed abundantly in the cell nucleus where it plays important roles in maintaining genomic integrity. The cleavage of PARP1 is a common marker for cell apoptosis pointing towards its breakdown in response to cellular stress.

Biological function summary

The enzymatic function of PARP1 involves the transfer of ADP-ribose units from NAD+ to target proteins a process known as ADP-ribosylation. PARP1 operates as a part of the base excision repair complex essential in DNA repair processes. The cleaved form of PARP1 no longer facilitates DNA repair marking a shift towards apoptosis. When PARP1 is cleaved it indicates caspase activity implying cells are undergoing programmed cell death.

Pathways

Cleaved PARP1 is deeply involved in the apoptosis and DNA damage response pathways. In the apoptosis pathway PARP1 interacts with key proteins like caspase-3 which cleaves PARP during apoptosis. In the DNA damage response PARP1 collaborates with proteins such as XRCC1 facilitating the base excision repair pathway important for fixing single-strand DNA breaks. These pathways highlight the dual role of PARP1 in promoting cell survival through repair and cell death via apoptosis.

Associated diseases and disorders

Cleaved PARP1 serves as an important marker in cancer and neurodegenerative diseases. In cancer research elevated levels of cleaved PARP1 suggest increased rates of apoptosis in response to anti-cancer therapies linking it to tumor suppression efforts. In neurodegenerative diseases excessive activation and cleavage of PARP1 can result in cell death exacerbating conditions like Alzheimer's disease. Through these contexts cleaved PARP1 connects to other therapeutic targets such as caspase proteins in cancer and to potential PARP inhibitors in neurodegenerative disorders.

Product promise

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11 product images

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Lanes 1 - 6: Merged signal (red and green). Green - ab32064 observed at 27 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab32064 was shown to react with Cleaved PARP1 in wild-type HAP1 cells in Western blot with loss of signal observed in PARP1 knockout sample.Wild-type HAP1 and PARP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab32064 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution

    Lane 1: Wild-type (1uM Staurosporine for 3hrs) HAP1 cell lysate at 20 µg

    Lane 2: Wild-type (Staurosporine control) HAP1 cell lysate at 20 µg

    Lane 3: PARP1 knockout (1uM Staurosporine for 3hrs) HAP1 cell lysate at 20 µg

    Lane 4: PARP1 knockout (Staurosporine control) HAP1 cell lysate at 20 µg

    Lane 5: HeLa (1uM Staurosporine for 3hrs) cell lysate at 20 µg

    Lane 6: HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 113 kDa

    Observed band size: 27 kDa

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Lane 1: Wild type HAP1 whole cell lysate (20 μg)
    Lane 2: PARP1 knockout HAP1 whole cell lysate (20 μg)
    Lane 3: HeLa whole cell lysate (20 μg)
    Lane 4: MCF7 whole cell lysate (20 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32064 observed at 30 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab32064 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. ab32064 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilutions.

    Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Predicted band size: 113 kDa

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Blocking/Dilution buffer 5% NFDM/TBST

    Exposure time :
    Lane 1,2: 1 second
    Lane 3,4: 8 seconds

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates treated with 1uM Staurosporine for 3 hours at 20 µg

    Lane 2: Untreated HeLa whole cell lysates at 20 µg

    Lane 3: NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysates treated with 1uM Staurosporine for 3 hours at 20 µg

    Lane 4: Untreated NIH/3T3 whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 113 kDa

    Observed band size: 25 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Immunohistochemical staining of paraffin embedded rat colon with purified ab32064 at a working dilution of 1/100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control (inset).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Immunohistochemical staining of paraffin embedded human ovarian carcinoma with purified ab32064 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. Counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control (inset).

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution

    Lane 1: Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg

    Lane 2: Jurkat cell lysate treated with camptothecin at 10 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 113 kDa

    Observed band size: 25 kDa

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

    Lane 1: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate at 20 µg

    Lane 2: NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 20 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 113 kDa

    Observed band size: 25 kDa

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Blockinng/Diluting buffer 5% NFDM/TBST

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/1000 dilution

    Lane 1: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysates treated with 1uM Staurosporine for 3 hours at 20 µg

    Lane 2: Untreated PC-12 whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 113 kDa

    Observed band size: 25 kDa

    Exposure time: 30s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Immunohistochemical staining of paraffin embedded human breast carcinoma tissue with unpurified ab32064 at a 1/100 dilution.

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Jurkat (Human T cell leukemia cell line from peripheral blood) cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of 15-Acetoxyscirpenol (ab142381). Increased expression of cleaved PARP1 (ab32064) in Jurkat cells correlates with an increase in 15-Acetoxyscirpenol concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab32064 at 1/10000 dilution and Anti-beta Actin antibody ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 and visualised using ECL development solution.

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 113 kDa

    Observed band size: 25 kDa

    Exposure time: 10s

  • Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064), expandable thumbnail

    Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064)

    Western blot: Anti-PARP1 antibody [E51] (ab32064) staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32064 was shown to bind specifically to PARP1. A band was observed at 27 kDa in wild-type A549 cell lysates with no signal observed at this size in PARP1 knockout cell line. To generate this image, wild-type and PARP1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution

    Lane 1: Wild-type A549 control staurosporine (0 uM, 72 h) cell lysate at 20 µg

    Lane 2: Wild-type A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

    Lane 3: Wild-type A549 control staurosporine (3 uM, 72 h) cell lysate at 20 µg

    Lane 4: PARP1 knockout A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

    Observed band size: 27 kDa

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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