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Synthetic peptide within Human Cleaved PARP1 aa 200-300. The exact sequence is proprietary. Residues following the cleavage of site.
This product is a recombinant rabbit monoclonal antibody.
A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
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.
|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.|
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).
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.
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.
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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"