Apoptosis Western Blot Cocktail (pro/p17-caspase 3, cleaved PARP1, muscle actin) (ab136812)

Overview

  • 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.

  • 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.

  • Tested applications
    Suitable for: WBmore details

Properties

Applications

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.

Application Abreviews Notes
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

Images



  • 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.

References

This product has been referenced in:
  • Tian L  et al. Siamese crocodile bile induces apoptosis in NCI-H1299 human non-small cell lung cancer cells via a mitochondria-mediated intrinsic pathway and inhibits tumorigenesis. Mol Med Rep 15:1727-1737 (2017). Read more (PubMed: 28259903) »
  • Wilson R  et al. MicroRNA regulation of endothelial TREX1 reprograms the tumour microenvironment. Nat Commun 7:13597 (2016). WB ; Human . Read more (PubMed: 27886180) »

See all 4 Publications for this product

Customer reviews and Q&As


We interpret the three FAS treatment specific bands in Jurkat cells as being various cleavage intermediates/products of the pro caspase 3 (uniprot P42574, http://www.uniprot.org/uniprot/P42574) processing. Specifically, amino acids 1-175 of P42574...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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