Recombinant 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), WB, ICC/IF, 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), WB, ICC/IF, 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/IF: 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...
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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 |
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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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WB |
1/1000. Predicted molecular weight: 85 kDa.
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ICC/IF | (1) |
Use at an assay dependent concentration.
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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. |
WB
1/1000. Predicted molecular weight: 85 kDa. |
ICC/IF
Use at an assay dependent concentration. |
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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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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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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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 -
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)This image is courtesy of an anonymous Abreview
ab32561 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.
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 (72)
ab32561 has been referenced in 72 publications.
- Takeuchi KI et al. Systemic innate immune response induces death of olfactory receptor neurons in Drosophila. Genes Cells 27:113-123 (2022). PubMed: 34921694
- Chen Q et al. Truncated PARP1 mediates ADP-ribosylation of RNA polymerase III for apoptosis. Cell Discov 8:3 (2022). PubMed: 35039483
- Zhu W et al. Long noncoding RNA SNHG8 promotes chemoresistance in gastric cancer via binding with hnRNPA1 and stabilizing TROY expression. Dig Liver Dis 54:1573-1582 (2022). PubMed: 35354542
- An W et al. Alpinia katsumadai Hayata induces growth inhibition and autophagy‑related apoptosis by regulating the AMPK and Akt/mTOR/p70S6K signaling pathways in cancer cells. Oncol Rep 48:N/A (2022). PubMed: 35730618
- Liu Z & Lv C RNA binding protein PUM2 promotes hepatocellular carcinoma proliferation and apoptosis via binding to the 3'UTR of BTG3. Oncol Lett 24:346 (2022). PubMed: 36072004