Recombinant Anti-Cleaved PARP1 antibody [SP276] (ab225715)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP276] to Cleaved PARP1
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cleaved PARP1 antibody [SP276]
See all Cleaved PARP1 primary antibodies -
Description
Rabbit monoclonal [SP276] to Cleaved PARP1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human tonsil and bladder transitional cell carcinoma tissues. WB: NIH/3T3 cell lysate treated with staurosporine.
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General notes
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA -
Concentration information loading...
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Purity
Protein A/G purified -
Purification notes
Purified from TCS by Protein A/G. -
Clonality
Monoclonal -
Clone number
SP276 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab225715 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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IHC-P |
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Primary incubation for 10 minutes at room temperature. |
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WB |
1/100. Predicted molecular weight: 113 kDa.
Primary incubation for 1 hour at room temperature. |
Notes |
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IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Primary incubation for 10 minutes at room temperature. |
WB
1/100. Predicted molecular weight: 113 kDa. Primary incubation for 1 hour at room temperature. |
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
- Entrez Gene: 11545 Mouse
- Entrez Gene: 25591 Rat
- Omim: 173870 Human
- SwissProt: P09874 Human
- SwissProt: P11103 Mouse
- SwissProt: P27008 Rat
- Unigene: 177766 Human
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 [SP276] (ab225715) at 1/100 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 : PARP1 knockout A549 control staurosporine (0 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: 113 kDa
Observed band size: 125,27 kDa why is the actual band size different from the predicted?Western blot: Anti-PARP1 antibody [SP276] (ab225715) staining at 1/100 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab225715 was shown to bind specifically to PARP1. A band was observed at 27/125 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 fluorescent western blot (TBS-based) blocking solution 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 [SP276] (ab225715) at 1/100 dilution
Lane 1 : Wild-type HAP1 Treated Staurosporine (1 uM, 3 h) cell lysate
Lane 2 : PARP1 knockout HAP1 Treated Staurosporine (1 uM, 3 h) cell lysate
Lane 3 : Wild-type HAP1 Control cell lysate
Lane 4 : PARP1 knockout HAP1 Control cell lysate
Lane 5 : HeLa Treated Staurosporine (1 uM, 3 h) cell lysate
Lane 6 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 113 kDa
Observed band size: 130, 27 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Cleaved PARP1 antibody [SP276] staining at 1/100 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab225715 was shown to bind specifically to Cleaved PARP1. A band was observed at 130 (full-length) and 27 (cleaved) kDa in treated wild-type HAP1 cell lysates with no signal observed at this size in PARP1 knockout cell line. To generate this image, wild-type and PARP1 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Formalin-fixed, paraffin-embedded human bladder transitional cell carcinoma tissue stained for Cleaved PARP1 using ab225715 at 1/100 dilution in immunohistochemical analysis.
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Formalin-fixed, paraffin-embedded human tonsil tissue stained for Cleaved PARP1 using ab225715 at 1/100 dilution in immunohistochemical analysis.
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All lanes : Anti-Cleaved PARP1 antibody [SP276] (ab225715) at 1/100 dilution
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) cell lysate treated with staurosporine
Lane 2 : Untreated NIH/3T3 (mouse embryo fibroblast cell line) cell lysate
Predicted band size: 113 kDa
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 (1)
ab225715 has been referenced in 1 publication.
- Tutuska K et al. Statin as anti-cancer therapy in autochthonous T-lymphomas expressing stabilized gain-of-function mutant p53 proteins. Cell Death Dis 11:274 (2020). PubMed: 32332697