Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E83-77] to Cleaved Caspase-3
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human, Recombinant fragment

Overview

  • Product name

    Anti-Cleaved Caspase-3 antibody [E83-77]
    See all Cleaved Caspase-3 primary antibodies

  • Description

    Rabbit monoclonal [E83-77] to Cleaved Caspase-3

  • Host species

    Rabbit

  • 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.

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Cleaved Caspase-3 aa 1-100 (N terminal). The exact sequence is proprietary. A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).
    Database link: P42574

  • Positive control

    • WB: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate; Jurkat cell lysate (camptothecin treated); HeLa cell lysate (staurosporine treated). ICC/IF: Hela cells (staurosporine treated); Human Vascular endothelial cells.

  • General notes

     

     

    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 production
    For more information see here.

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

Properties

Images

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    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 (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 (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 (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, 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 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    Cells were grown to confluency prior to treatment.

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

    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.

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

    Ab32042, at dilution of 1/100, staining HeLa (human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.
    Left image: control.
    Right image: staurosporine treated.

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    All lanes : 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
    Additional bands at: 17 kDa (possible mature (processed) protein)

  • Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
    Lane 1 : anti Pro Caspase 3 at 1/10000 dilution
    Lane 2 : anti Pro Caspase 3 at 1/10000 dilution
    Lanes 3-4 : 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 & 4 : Jurkat cell lysate + Camptothecin
    Lane 3 : Jurkat cell lysate

    Predicted band size: 32 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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