Product namep53 Mediated Apoptosis WB Cocktail
Species reactivityReacts with: Human
The p53-Mediated Apoptosis Cocktail is designed to study the induction of p53-mediated apoptosis in response to various stimuli. The two main components of this cocktail are monoclonal antibodies specific to PARP (cleaved) and p53 (phosphorylated at serine 46). The GAPDH antibody is provided as a loading control for sample to sample normalization. Since the primary antibodies are both mouse and rabbit, the cocktail of HRP-conjugated goat anti-rabbit and anti-mouse secondary antibodies is provided for convenience. The targets are easily resolved by Western blot given their different molecular weights.
Poly [ADP-ribose] polymerase 1 (PARP) is a DNA repair enzyme that is cleaved during apoptosis by activated caspases. The mouse PARP antibody of this cocktail detects only the apoptosis-specific 89 kDa PARP fragment (cleaved-PARP). This antibody does not react with the full-length PARP. The tumor suppressor protein p53 is an important component in the regulation of cellular growth. p53 regulates the cellular response to DNA damage and other forms of cellular stress by inducing activation of genes involved in DNA repair, cell cycle control, and apoptosis. Phosphorylation of p53 at serine 46 is a critical part of p53-mediated apoptosis, allowing for the induction of various pro-apoptotic genes.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at +4°C. Please refer to protocols.
Components 200 µl 100X HRP Conjugated Secondary Antibody Cocktail 1 x 500µl 250X p53-Mediated Apoptosis WB Cocktail 1 x 200µl
Our Abpromise guarantee covers the use of ab140360 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.
Positive control is MCF7 cells treated with 1 uM camptothecin for 16 hours.
Suggested working concentration 1/250 dilution for primary antibodies and 1/100 dilution for secondary antibodies.
Lane 1: MCF7, vehicle (DMSO) treated Lane 2: MCF7, 1 µM camptothecin for 6 hours Lane 3: MCF7, 1 µM camptothecin for 16 hours Lane 4: MCF7, 1 µM camptothecin for 24 hours Primary antibodies All lanes: ab140360 primary antibodies cocktail at a 1/250 dilution Secondary antibodies All lanes: ab140360 HRP, GAR-HRP cocktail at a 1/100 dilution
Lanes 1, 3, 5, 7: MCF7: vehicle (DMSO) treated for 16 hours Lanes 2, 4, 6, 8: MCF7: 1 µM camptothecin for 16 hours All lysates at 20 µg per lane. Primary antibodies Lanes 1, 2: cleaved PARP Lanes 3, 4: p53 pSer46 Lanes 5, 6: GAPDH Lanes 7, 8: ab140360 primary antibodies cocktail at a 1/250 dilution Secondary antibodies All lanes: ab140360 GAM-HRP, GAR-HRP cocktail at a 1/100 dilution