Recombinant Anti-Cleaved PARP1 antibody [E51] (ab32064)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E51] to Cleaved PARP1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cleaved PARP1 antibody [E51]
See all Cleaved PARP1 primary antibodies -
Description
Rabbit monoclonal [E51] to Cleaved PARP1 -
Host species
Rabbit -
Specificity
This antibody is specific for the p25 cleaved form of human PARP1. -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chinese hamster -
Immunogen
Synthetic peptide within Human Cleaved PARP1 aa 150-250. The exact sequence is proprietary.
Database link: P09874 -
Positive control
- WB: Jurkat whole cell lysate (ab7899). HeLa and RAW 264.7 whole cell lysate. HAP1, HeLa, NIH/3T3 and PC-12 treated with 1uM Staurosporine. Jukat cells treated with camptothecin. Jukat cells treated with 15-Acetoxyscirpenol. IHC-P: Rat colon tissue. Human ovarian cancer and breast carcinoma tissue.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E51 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32064 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (7) |
1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
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IHC-P | (5) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 25 kDa. |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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. 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 similarities
Contains 1 BRCT domain.
Contains 1 PARP alpha-helical domain.
Contains 1 PARP catalytic domain.
Contains 2 PARP-type zinc fingers. -
Post-translational
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 localization
Nucleus. Nucleus, nucleolus. Localizes at sites of DNA damage. - Information by UniProt
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Database links
- Entrez Gene: 100689463 Chinese hamster
- Entrez Gene: 142 Human
- Entrez Gene: 11545 Mouse
- Entrez Gene: 25591 Rat
- Omim: 173870 Human
- SwissProt: Q9R152 Chinese hamster
- SwissProt: P09874 Human
- SwissProt: P11103 Mouse
see all -
Alternative names
- ADP ribosyltransferase diphtheria toxin like 1 antibody
- ADP ribosyltransferase NAD(+) antibody
- ADP-ribosyltransferase diphtheria toxin-like 1 antibody
see all
Images
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All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution
Lane 1 : Wild-type A549 control staurosporine (0 uM, 72 h) cell lysate
Lane 2 : Wild-type A549 treated staurosporine (3 uM, 24 h) cell lysate
Lane 3 : Wild-type A549 control staurosporine (3 uM, 72 h) cell lysate
Lane 4 : PARP1 knockout A549 treated staurosporine (3 uM, 24 h) cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?Western blot: Anti-PARP1 antibody [E51] (ab32064) staining at 1/10000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (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.
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All lanes : Anti-Cleaved PARP1 antibody [E51] (ab32064) at 1/10000 dilution
Lane 1 : Wild-type (1uM Staurosporine for 3hrs) HAP1 cell lysate
Lane 2 : Wild-type (Staurosporine control) HAP1 cell lysate
Lane 3 : PARP1 knockout (1uM Staurosporine for 3hrs) HAP1 cell lysate
Lane 4 : PARP1 knockout (Staurosporine control) HAP1 cell lysate
Lane 5 : HeLa (1uM Staurosporine for 3hrs) cell lysate
Lane 6 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab32064 observed at 27 kDa. Red - 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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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).
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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, 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 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) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : 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
Lane 2 : Untreated HeLa whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysates treated with 1uM Staurosporine for 3 hours
Lane 4 : Untreated NIH/3T3 whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking/Dilution buffer 5% NFDM/TBST
Exposure time :
Lane 1,2: 1 second
Lane 3,4: 8 seconds -
Immunohistochemical staining of paraffin embedded human breast carcinoma tissue with unpurified ab32064 at a 1/100 dilution.
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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).
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All lanes : 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
Lane 2 : Jurkat cell lysate treated with camptothecin
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : 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
Lane 2 : NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : 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
Lane 2 : Untreated PC-12 whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 25 kDa
Exposure time: 30 secondsBlockinng/Diluting buffer 5% NFDM/TBST
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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 ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (344)
ab32064 has been referenced in 344 publications.
- Li J et al. METTL3 promotes prostatic hyperplasia by regulating PTEN expression in an m6A-YTHDF2-dependent manner. Cell Death Dis 13:723 (2022). WB ; Human . PubMed: 35985997
- Wu G et al. CircASPH promotes KGN cells proliferation through miR-375/MAP2K6 axis in Polycystic Ovary Syndrome. J Cell Mol Med 26:1817-1825 (2022). PubMed: 33372369
- Felip E et al. Modulation of DNA Damage Response by SAM and HD Domain Containing Deoxynucleoside Triphosphate Triphosphohydrolase (SAMHD1) Determines Prognosis and Treatment Efficacy in Different Solid Tumor Types. Cancers (Basel) 14:N/A (2022). PubMed: 35158911
- Knoell J et al. PACS2-TRPV1 axis is required for ER-mitochondrial tethering during ER stress and lung fibrosis. Cell Mol Life Sci 79:151 (2022). PubMed: 35212819
- Shi L et al. miR-146b-5p promotes colorectal cancer progression by targeting TRAF6. Exp Ther Med 23:231 (2022). PubMed: 35222708