Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Caspase-9 antibody [E84] (ab32068)

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Overview

  • Product name

    Anti-Caspase-9 antibody [E84]
    See all Caspase-9 primary antibodies

  • Description

    Rabbit monoclonal [E84] to Caspase-9

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-Pmore details
    Unsuitable for: ICC or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Caspase-9 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • Jurkat cell lysate

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32068 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 1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P 1/25. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC or IP.

    • Target

    • Function

      Involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP).
      Isoform 2 lacks activity is an dominant-negative inhibitor of caspase-9.

    • Tissue specificity

      Ubiquitous, with highest expression in the heart, moderate expression in liver, skeletal muscle, and pancreas. Low levels in all other tissues. Within the heart, specifically expressed in myocytes.

    • Sequence similarities

      Belongs to the peptidase C14A family.
      Contains 1 CARD domain.

    • Developmental stage

      Expressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart.

    • Post-translational
      modifications

      Cleavages at Asp-315 by granzyme B and at Asp-330 by caspase-3 generate the two active subunits. Caspase-8 and -10 can also be involved in these processing events.
    • Information by UniProt

    • Database links

      • Alternative names

        • APAF-3 antibody
        • APAF3 antibody
        • Apoptosis related cysteine peptidase antibody
        • Apoptotic protease Mch-6 antibody
        • Apoptotic protease-activating factor 3 antibody
        • CASP-9 antibody
        • CASP9 antibody
        • CASP9_HUMAN antibody
        • Caspase 9 apoptosis related cysteine peptidase antibody
        • Caspase 9 Dominant Negative antibody
        • Caspase 9c antibody
        • Caspase-9 antibody
        • Caspase-9 subunit p10 antibody
        • ICE LAP6 antibody
        • ICE like apoptotic protease 6 antibody
        • ICE-LAP6 antibody
        • ICE-like apoptotic protease 6 antibody
        • MCH6 antibody
        • PPP1R56 antibody
        • protein phosphatase 1, regulatory subunit 56 antibody
        • RNCASP9 antibody
        see all

      Images

      • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
        Western blot - Anti-Caspase-9 antibody [E84] (ab32068)

        Lanes:
        Lane 1: Wild-type HAP1 whole cell lysate (40 µg).
        Lane 2: CASP9 knockout HAP1 whole cell lysate (40µg).
        Lane 3: HeLa whole cell lysate (40 µg).

        Lanes 1 - 3 Merged signal (red and green). Green - ab32068 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.

        ab32068 was shown to specifically recognize CASP9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CASP9 knockout samples were examined. Wild-type and CASP9 knockout samples were subjected to SDS-PAGE. Ab32068 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
        Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
        All lanes : Anti-Caspase-9 antibody [E84] (ab32068) at 1/1000 dilution

        Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
        Lane 2 : HeLa treated with staurosporine 1uM for 4 hours whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 46 kDa
        Additional bands at: 35 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


        Exposure time: 3 minutes


        Blocking/dilution buffer: 5% NFDM/TBST.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)

        Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-Pro-Caspase-9 RabMAb (ab32068).

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
        Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
        Anti-Caspase-9 antibody [E84] (ab32068) at 1/2000 dilution + Jurkat cell lysate at 10 µg

        Predicted band size: 46 kDa
        Observed band size: 46 kDa

      References

      This product has been referenced in:

      • Gu LL  et al. Oxymatrine Causes Hepatotoxicity by Promoting the Phosphorylation of JNK and Induction of Endoplasmic Reticulum Stress Mediated by ROS in LO2 Cells. Mol Cells 41:401-412 (2018). Read more (PubMed: 29754474) »
      • Liu Y & Zhang Y Lycium barbarum polysaccharides alleviate hydrogen peroxide-induced injury by up-regulation of miR-4295 in human trabecular meshwork cells. Exp Mol Pathol 106:109-115 (2018). Read more (PubMed: 30594603) »
      See all 8 Publications for this product

      Publishing research using ab32068? Please let us know so that we can cite the reference in this datasheet.

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