Product nameApoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin)
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
Cocktail of primary antibodies to detect apoptosis biomarkers caspase 3 and PARP, along with loading control muscle actin (42 kDa). The caspase 3 antibody (rabbit monoclonal) detects both the 32 kDa pro-caspase 3 as well as the p17 subunit of active caspase 3 generated by cleavage of the pro-caspase 3 at Asp175. The PARP antibody (mouse monoclonal) detects only the apoptosis-specific 89 kDa PARP fragment (cleaved-PARP) generated from the full length PARP by active caspases. Since the primary antibodies used are both mouse and rabbit, a secondary antibodies cocktail of GAM-HRP and GAR-HRP is provided.
The Apoptosis western blot cocktail (ab136812) is designed to study the induction of apoptosis in response to various stimuli. The two main components of this cocktail are monoclonal antibodies specific to caspase 3 and PARP. Caspase 3 is one of the executioner caspases activated by proteolytic cleavage during apoptosis. The rabbit caspase 3 antibody of this cocktail detects both the 32 kDa pro-caspase 3 as well as the p17 subunit of the active caspase 3 generated by cleavage of the pro-caspase 3 at Asp175. Thus the induction of apoptosis can be followed by a decrease of the pro-caspase 3 or by an increase of the p17 caspase 3. Monitoring the changes in the pro-caspase 3 is particularly advantageous, since the proportion of caspase activation can be determined from the reduction of the pro-form from analysis of control and stimulated samples. 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. Combined, these two antibodies provide biomarkers of apoptosis. The rabbit muscle actin 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.
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 Primary Antibody Cocktail 1 x 200µl
- Actin, alpha skeletal muscle
- ADP-ribosyltransferase diphtheria toxin-like 1
Our Abpromise guarantee covers the use of ab136812 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.
1/250 dilution for primary antibodies
1/100 dilution for secondary antibodies
Suggested dilution buffer: 5% milk/PBS+0.05% Tween 20
Lane 1: Jurkat cells, untreated
Lane 2: Jurkat cells treated with anti-FAS for 2 hours
Lane 3: Jurkat cells treated with anti-FAS for 4 hours
Lane 4: Jurkat cells treated with anti-FAS for 6 hours
All lysates at 20 µg/lane
All lanes: 250X Primary Antibodies Cocktail, 1/250 dilution.
All lanes: 100X HRP-Conjugated Secondary Antibodies Cocktail (ab136812), 1/100 dilution.
Lanes 1, 3, 5, 7: (ab136806) HeLa, vehicle treated
Lanes 2, 4, 6, 8: (ab136806) HeLa, 1 µM staurosporine (ab120056), 4 hours
All lysates at 20 µg per lane.
Lanes 1, 2: Cleaved PARP
Lanes 3, 4: Actin
Lanes 5, 6: Caspase 3
Lanes 7, 8: ab136812 250X Primary Antibodies Cocktail, 1/250 dilution
All lanes: ab136812 100X HRP-Conjugated Secondary Antibodies Cocktail, 1/100 dilution.
This product has been referenced in:
- Deben C et al. Hypoxia-Induced Cisplatin Resistance in Non-Small Cell Lung Cancer Cells Is Mediated by HIF-1a and Mutant p53 and Can Be Overcome by Induction of Oxidative Stress. Cancers (Basel) 10:N/A (2018). Read more (PubMed: 29690507) »
- Bhuyan DJ et al. Eucalyptus microcorys leaf extract derived HPLC-fraction reduces the viability of MIA PaCa-2 cells by inducing apoptosis and arresting cell cycle. Biomed Pharmacother 105:449-460 (2018). WB . Read more (PubMed: 29879629) »