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Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human active Caspase-3 aa 1-100 (N terminal). A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).
Database link: P42574
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.
Our Abpromise guarantee covers the use of ab32042 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).|
|ICC/IF||1/100 - 1/250.|
Lane 1: Wild type HAP1 + DMSO, 24 hours whole cell lysate (20 µg)
Lane 2: HAP1 + 2uM Staurosporine (ab146588), 24 hours knockout HAP1 + DMSO, 24 hours whole cell lysate (20 µg)
Lane 3: CASP3 KO + EDTA, 24 hours whole cell lysate (20 µg)
Lane 4: CASP3 KO + 2uM Staurosporine (ab146588), 24 hours whole cell lysate (20 µg)
Lane 5: HeLa + EDTA, 24 hours whole cell lysate (20 µg)
Lane 6: HeLa + 2uM Staurosporine (ab146588), 24 hours whole cell lysate (20 µg)
Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, ab8245, observed at 130 kDa.
ab32042 was shown to specifically react with HAP1 + 2uM Staurosporine (ab146588), 24 hours when HAP1 + 2uM Staurosporine (ab146588) 24 hours knockout samples were used. Wild-type and HAP1 + 2uM Staurosporine (ab146588), 24 hours knockout samples were subjected to SDS-PAGE. Ab32042 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 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.
Cells were grown to confluency prior to treatment.
Immunohistochemical analysis of FFPE Human prostate cancer tissue, labeling active Caspase-3 with ab32042. Samples were fixed in formaldehyde, antigen retrieval was by heat mediated treatment with 0.05% Trysin, permeabilization was with 0.1% Triton X-100 and blocking with 10% serum for 1 hour at 23°C. Immunostaining with anti-active Caspase-3 diluted 1/200 for 16 hours at 4°C.