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Anti-Cleaved Caspase-3 antibody [E83-77] ab32042 is a rabbit monoclonal antibody that is used in Cleaved Caspase-3 western blotting and immunofluorescence. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone E83-77 has been tried and trusted by researchers since 2006 and is cited in >860 publications
- Specificity confirmed with CASP3 knockout cell line validation
New 20 ul size available


Images

Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail
  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail
  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail
  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail
  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (AB32042), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow CytIPWBIHC-PICC/IF
Human
Not recommended
Not recommended
Tested
Not recommended
Tested
Recombinant fragment
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Recombinant fragment - Human
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Not recommended
Not recommended

Species

Recombinant fragment, Human, Recombinant fragment - Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Recombinant fragment, Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant fragment - Human

Dilution info

1/500

Notes

The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.

Species

Human

Dilution info

1/500

Notes

The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.

Not recommended
Not recommended

Species

Recombinant fragment

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Recombinant fragment, Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment, Recombinant fragment - Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

4 products for Alternative Product

Target data

Function

Involved in the activation cascade of caspases responsible for apoptosis execution (PubMed:7596430). At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-|-Gly-217' bond (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. Cleaves and activates caspase-6, -7 and -9 (PubMed:7596430). 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).

Alternative names

Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1, CASP3, CPP32

Recommended products

Anti-Cleaved Caspase-3 antibody [E83-77] ab32042 is a rabbit monoclonal antibody that is used in Cleaved Caspase-3 western blotting and immunofluorescence. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone E83-77 has been tried and trusted by researchers since 2006 and is cited in >860 publications
- Specificity confirmed with CASP3 knockout cell line validation
New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

E83-77

Purification technique

Affinity purification Protein A

Specificity

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information. This antibody only detects the active (cleaved) form of Caspase-3 and does not recognize the pro form of Caspase-3. PubMedID 19789217 describes the detection of human cells injected into mice. The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Cleaved Caspase-3 also known as active caspase 3 plays an important role in the execution phase of cell apoptosis. The caspase 3 molecule has a molecular weight of approximately 32 kDa. It undergoes cleavage into subunits that are important for its activation marking its conversion into the cleaved form. Cleaved caspase is widely expressed in various tissues with a notable presence in the cytoplasm of cells undergoing apoptotic processes. Cleavage of caspase 3 results in the formation of a functional enzyme that participates in the degradation of cellular components during apoptosis.

Biological function summary

Caspase 3 acts as an executioner enzyme in the process of programmed cell death. Its cleavage from the inactive zymogen form into the active form triggers apoptotic pathways ensuring the removal of damaged or unnecessary cells. Cleaved caspase operates within a proteolytic cascade often in conjunction with other caspases such as caspase 9. Once activated caspase 3 targets and cleaves specific cellular substrates contributing to the characteristic morphological changes and DNA fragmentation associated with apoptosis.

Pathways

Cleaved caspase 3 is significant within the intrinsic and extrinsic apoptotic pathways. It acts downstream of initiator caspases such as caspase 9 in the intrinsic pathway where its activation involves mitochondrial signals. In the extrinsic pathway it closely associates with caspase 8 mediating cell death signals from extracellular triggers. These pathways ensure that cleaved caspase 3 fulfills its role as a critical mediator of apoptosis linking upstream death signals to cellular dismantling during the apoptosis process.

Associated diseases and disorders

Cleaved caspase 3 connects to conditions such as cancer and neurodegenerative diseases. In cancer dysregulation of caspase 3 activity may lead to evasion of apoptosis facilitating tumor progression. Therapeutic approaches aim to restore its apoptotic functions in cancer cells. In neurodegenerative diseases like Alzheimer's disease abnormal activation of cleaved caspase 3 contributes to neuronal loss. It interacts with proteins like amyloid precursor protein promoting the pathological changes associated with this disorder. Understanding these interactions provides insights into potential therapeutic avenues.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail
    Image from Zhu ZX et al., PLoS One. 2015;10(12):e0144495. Fig 7.; doi: 10.1371/journal.pone.0144495. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    High-glucose induces apoptosis in human Vascular endothelial cells (VECs).

    Apoptotic responses in VEC were analyzed by detection of cleaved-Caspase-3 immunofluorescence using ab32042. Cells were treated with low glucose (LG) or high glucose (HG) for 72 hours before treated with 100 ng/mL bFGF, 1 μM sp600125 (sp), or 1 μM U0126 (U) or 10 μM MnTmPyP for 1 hour. Bar = 100 μm.

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail

    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    Lane 1: Wild type HAP1 + DMSO for 24 hours, whole cell lysate (20 μg)
    Lane 2: Wild type HAP1 + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
    Lane 3: HAP1 CASP3 KO + DMSO for 24 hours, whole cell lysate (20 μg)
    Lane 4: HAP1 CASP3 KO + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
    Lane 5: HeLa + DMSO for 24 hours, whole cell lysate (20 μg)
    Lane 6: HeLa + 2uM Staurosporine (Staurosporine (DMSO solution), protein kinase inhibitor ab146588) for 24 hours, whole cell lysate (20 μg)
    Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.

    ab32042 was shown to specifically react with CASP3 (Caspase-3) when CASP3 (Caspase-3) knockout samples were used. HAP1 wild-type and CASP3 (Caspase-3) knockout samples were subjected to SDS-PAGE. ab32042 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    Cells were grown to confluency prior to treatment.

    All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    Predicted band size: 32 kDa

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail

    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution

    Lane 1: HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg

    Lane 2: HeLa Whole Cell Lysate (untreated) at 20 µg

    Lane 3: Cleaved Caspase 3 (recombinant protein) at 0.1 µg

    Secondary

    All lanes: 800CW Goat Anti-Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 32 kDa

    Observed band size: 17 kDa

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail

    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    Lanes 1 - 2: anti Pro Caspase 3 at 1/10000 dilution

    Lanes 3 - 4: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution

    Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate

    Lanes 2 and 4: Jurkat cell lysate + Camptothecin

    Lane 3: Jurkat cell lysate

    Predicted band size: 32 kDa

    Observed band size: 17 kDa, 30 kDa

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042), expandable thumbnail

    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)

    Western blot: Anti-CASP3 antibody [E83-77] (ab32042) staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32042 was shown to bind specifically to CASP3. A band was observed at 16/28 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in CASP3 knockout cell line. To generate this image, wild-type and CASP3 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution

    Lane 1: Wild-type HeLa Treated Staurosporine (2uM, 4h) cell lysate at 20 µg

    Lane 2: CASP3 knockout HeLa Treated Staurosporine (2uM, 4h) cell lysate at 20 µg

    Lane 3: Wild-type HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate, at 20 µg

    Lane 4: CASP3 knockout HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 32 kDa

    Observed band size: 16 kDa, 28 kDa

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Product protocols

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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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