Recombinant Anti-Cleaved PARP1 antibody [Y34] (ab32561)
![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)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y34] to Cleaved PARP1
- Suitable for: Flow Cyt (Intra), ICC, WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
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: Flow Cyt (Intra), ICC, WB, IPmore 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). IP: HeLa cell lysate. ICC: HeLa cells
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
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
The Abpromise guarantee
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 |
|---|---|---|
| Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC |
Use at an assay dependent concentration.
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| WB |
1/1000. Predicted molecular weight: 85 kDa.
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| IP |
1/50.
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| Notes |
|---|
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC
Use at an assay dependent concentration. |
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WB
1/1000. Predicted molecular weight: 85 kDa. |
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IP
1/50. |
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)
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 (Intracellular) - 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 (Intracellular) - Anti-Cleaved PARP1 antibody [Y34] (ab32561)Primary ab 1/50 dilution (0.5µg / Red). Secondary abGoat 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 Intracellular 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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![Immunoprecipitation - Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-7-anti-cleaved-parp1-antibody-y34-immunoprecipitation-hela-human.jpg)
Immunoprecipitation - Anti-Cleaved PARP1 antibody [Y34] (ab32561)Purified ab32561 at 1/50 dilution (2µg) immunoprecipitating Cleaved PARP1 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32561 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32561 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 85 kDa -
![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 -
![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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![Anti-Cleaved PARP1 antibody [Y34] (ab32561)](https://www.abcam.com/ps/products/32/ab32561/Images/ab32561-6-benefits-of-recombinant-antibodies.png)
Anti-Cleaved PARP1 antibody [Y34] (ab32561)
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 (60)
ab32561 has been referenced in 60 publications.
- Shi C et al. lncRNA SNHG14 Plays a Role in Sepsis-Induced Acute Kidney Injury by Regulating miR-93. Mediators Inflamm 2021:5318369 (2021). PubMed: 33505213
- Wang L et al. miR-4478 sensitizes ovarian cancer cells to irradiation by inhibiting Fus and attenuating autophagy. Mol Ther Nucleic Acids 23:1110-1119 (2021). PubMed: 33664992
- Liebl MC et al. DAZAP2 acts as specifier of the p53 response to DNA damage. Nucleic Acids Res 49:2759-2776 (2021). PubMed: 33591310
- Ma YS et al. MicroRNA-499 serves as a sensitizer for lung cancer cells to radiotherapy by inhibition of CK2a-mediated phosphorylation of p65. Mol Ther Oncolytics 21:171-182 (2021). PubMed: 33997273
- Li X et al. Circ0120816 acts as an oncogene of esophageal squamous cell carcinoma by inhibiting miR-1305 and releasing TXNRD1. Cancer Cell Int 20:526 (2020). PubMed: 33292234