Recombinant Anti-Cleaved PARP1 antibody [Y34] (ab32561)
Overview
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Product name
Anti-Cleaved PARP1 antibody [Y34]
See all Cleaved PARP1 primary antibodies -
Description
Rabbit monoclonal [Y34] to Cleaved PARP1 -
Host species
Rabbit -
Specificity
This antibody is specific for p85 cleaved form of PARP1. -
Tested applications
Suitable for: IHC-P, WB, Flow Cyt, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Cleaved PARP1 aa 200-300. The exact sequence is proprietary. Residues following the cleavage of site.
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Positive control
- Jurkat whole cell lysate (ab7899).
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
IgG fraction -
Clonality
Monoclonal -
Clone number
Y34 -
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
Applications
Our Abpromise guarantee covers the use of ab32561 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 |
|---|---|---|
| IHC-P | Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. PubMed: 21931707 | |
| WB | 1/1000. Predicted molecular weight: 85 kDa. | |
| Flow Cyt | 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IP | 1/50. | |
| ICC/IF | 1/500. |
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: 142 Human
- Omim: 173870 Human
- SwissProt: P09874 Human
- Unigene: 177766 Human
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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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![Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-278220-anti-cleaved-parp1-antibody-y34-western-blot.jpg)
Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)Lane 1: Wild type HAP1 (untreated) whole cell lysate (20 µg)
Lane 2: PARP1 (untreated) knockout HAP1 (untreated) whole cell lysate (20 µg)
Lane 3: HeLa (untreated) whole cell lysate (20 µg)
Lane 4: HAP1 (staurosporin treated, 1 u M, 4 hr) whole cell lysate (20 µg)
Lane 5: PARP1 (staurosporin treated, 1 uM, 4 hr) knockout HAP1 whole cell lysate (20 µg)
Lane 6: HeLa (staurosporin treated, 1 uM, 4 hr) whole cell lysate (20 µg)Lanes 1 - 6: Merged signal (red and green). Green - ab32561 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32561 was shown to specifically react with PARP1 (untreated) when PARP1 (untreated) knockout samples were used. Wild-type and PARP1 (untreated) knockout samples were subjected to SDS-PAGE. Ab32561 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.
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![Flow Cytometry - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-280634-anti-cleaved-parp1-antibody-y34-flow-cytometry.jpg)
Flow Cytometry - Anti-Cleaved PARP1 antibody [Y34] (ab32561)Primary ab 1/50 dilution (0.5μg / Red). Secondary ab Goat anti rabbit IgG (FITC). Secondary ab concentration 1/150 dilution. Cell line Jurkat (human acute T cell leukemia) treated with (Right) or without (Left) 4μM Camptothecin for 5h. Fixative 4% paraformaldehyde. Datasheet comment Flow cytometric analysis of apoptotic and non-apoptotic Jurkat cells using anti-cleaved PARP1 RabMAb (ab32561). Jurkat cells were either left untreated (A) or treated with camptothecin (4 uM, 5 hr) to induce apoptosis (B). Cells were fixed and permeabilized, and then stained with anti-cleaved PARP1. The results indicate that 43% of cells were positive for cleaved PARP1 (B, M2) after treatment, compared to 9% positive without treatment (A, M2).
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![Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-237748-anti-cleaved-parp1-antibody-y34-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP1 antibody [Y34] (ab32561)This image is courtesy of an anonymous Abreviewab32561 staining Cleaved PARP1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.
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![Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-45258-anti-cleaved-parp1-antibody-y34-western-blot.jpg)
Western blot - Anti-Cleaved PARP1 antibody [Y34] (ab32561)All lanes : Anti-Cleaved PARP1 antibody [Y34] (ab32561) at 1/1000 dilution
Lane 1 : Un-treated Jurkat cell lysate.
Lane 2 : Jurkat cell lysate treated with Camptothecin.
Predicted band size: 85 kDa
Observed band size: 85 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-45257-anti-cleaved-parp1-antibody-y34-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)Image from Wang L et al., PLoS One. 2011;6(9):e24405. Epub 2011 Sep 9. Fig 6.; doi:10.1371/journal.pone.0024405; September 9, 2011, PLoS ONE 6(9): e24405.Immunohistochemical analysis of OVCAR-3 tumour xenografts in nude mice, staining cleaved PARP1 with ab32561.
Antigen retrieval was performed via heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100) overnight at 4°C. A biotinyated anti-rabbit IgG (1/150) was used as the secondary antibody and staining was detected using DAB.
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.
References
ab32561 has been referenced in 33 publications.
- Chen Y et al. Epigenetically upregulated oncoprotein PLCE1 drives esophageal carcinoma angiogenesis and proliferation via activating the PI-PLCe-NF-?B signaling pathway and VEGF-C/ Bcl-2 expression. Mol Cancer 18:1 (2019). PubMed: 30609930
- Ma C et al. Emodin induces apoptosis and autophagy of fibroblasts obtained from patient with ankylosing spondylitis. Drug Des Devel Ther 13:601-609 (2019). PubMed: 30809091
- Sun L et al. Downregulation of ENDOCAN in myeloid leukemia cells inhibits proliferation and promotes apoptosis by suppressing nuclear factor-?B activity. Mol Med Rep 19:3247-3254 (2019). PubMed: 30816462
- Wu CE et al. Cinnamaldehyde enhances apoptotic effect of oxaliplatin and reverses epithelial-mesenchymal transition and stemnness in hypoxic colorectal cancer cells. Exp Cell Res 383:111500 (2019). PubMed: 31306656
- Wu F et al. HOXA6 inhibits cell proliferation and induces apoptosis by suppressing the PI3K/Akt signaling pathway in clear cell renal cell carcinoma. Int J Oncol 54:2095-2105 (2019). PubMed: 31081053
