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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.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
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 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"