Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) (ab136812)
Overview
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Product name
Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Product overview
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.
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Notes
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.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store 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 -
Research areas
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Alternative names
- ACTA1
- Actin, alpha skeletal muscle
- ADP-ribosyltransferase diphtheria toxin-like 1
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Database links
- Entrez Gene: 58 Human
- Entrez Gene: 142 Human
- Entrez Gene: 836 Human
- Omim: 173870 Human
- Omim: 102610 Human
- Omim: 600636 Human
- SwissProt: P42574 Human
- SwissProt: P09874 Human
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Images
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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
Primary antibodies
All lanes: 250X Primary Antibodies Cocktail, 1/250 dilution.
Secondary antibodies
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.
Primary antibodies
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
Secondary antibodies
All lanes: ab136812 100X HRP-Conjugated Secondary Antibodies Cocktail, 1/100 dilution.