The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Predicted molecular weight: 85 kDa.
1/50. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Application notesIs unsuitable for IHC.
FunctionInvolved 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.
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 localizationNucleus. Nucleus, nucleolus. Localizes at sites of DNA damage.
Western blot - Anti-Cleaved PARP antibody [Y34] (ab32561)
Predicted band size : 85 kDa
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) 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.
Immunocytochemistry/ Immunofluorescence - Anti-Cleaved PARP antibody [Y34] (ab32561)This image is courtesy of an anonymous Abreview
ab32561 staining Cleaved PARP 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.
Western blot - cleaved PARP antibody [Y34] (ab32561)
All lanes : Anti-Cleaved PARP 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 PARP 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 PARP 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.
References for Anti-Cleaved PARP antibody [Y34] (ab32561)
This product has been referenced in:
Peh J et al. The Combination of Vemurafenib and Procaspase-3 Activation Is Synergistic in Mutant BRAF Melanomas. Mol Cancer Ther15:1859-69 (2016).
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