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Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) (ab136812) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.

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Images

Western blot - Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) (AB136812), expandable thumbnail
  • Western blot - Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) (AB136812), expandable thumbnail

Publications

Key facts

Applications

WB

Target

CASP3
(See target data below)

Reactive species

Human

Reactivity data

Application

WB

Reactivity

Reacts

Dilution info

-

Notes

1/250 dilution for primary antibodies. 1/100 dilution for secondary antibodies. Suggested dilution buffer: 5% milk/PBS+0.05% Tween 20.

Target data

Function

Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:35338844, PubMed:35446120, PubMed:7596430). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:7596430). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond (PubMed:10497198, PubMed:16374543, PubMed:7596430, PubMed:7774019). Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain (By similarity). Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively) (PubMed:7596430). Cleaves and inactivates interleukin-18 (IL18) (PubMed:37993714, PubMed:9334240). Involved in the cleavage of huntingtin (PubMed:8696339). Triggers cell adhesion in sympathetic neurons through RET cleavage (PubMed:21357690). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:23152800). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (PubMed:35338844, PubMed:35446120). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:23845944, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758104, PubMed:36758106).

Targets

PARP1, ACTC1

Alternative names

Recommended products

Apoptosis Western Blot Cocktail (pro/p17-caspase-3, cleaved PARP1, muscle actin) (ab136812) is part of the reagents, controls & accessories range. Abcam offers high-quality biological reagents and tools including antibodies, proteins, assays, cell lines and lysates.

Key facts

Applications

WB

Target

CASP3

Reactive species

Human

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Notes

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.

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2 product images

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

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




    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.

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

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


    Lanes 1, 3, 5, 7: (HeLa Apoptosis Lysate Set: Staurosporine-Treated and Vehicle-Treated Control ab136806) HeLa, vehicle treated

    Lanes 2, 4, 6, 8: (HeLa Apoptosis Lysate Set: Staurosporine-Treated and Vehicle-Treated Control ab136806) HeLa, 1 µM staurosporine (Staurosporine, Protein kinase inhibitor 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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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