The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Application notesIs unsuitable for ICC or IP.
FunctionInvolved 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 specificityUbiquitous, 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 similaritiesBelongs to the peptidase C14A family. Contains 1 CARD domain.
Developmental stageExpressed at low levels in fetal heart, at moderate levels in neonate heart, and at high levels in adult heart.
Post-translational modificationsCleavages 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. Phosphorylated at Thr-125 by MAPK1/ERK2. Phosphorylation at Thr-125 is sufficient to block caspase-9 processing and subsequent caspase-3 activation.
Western blot - Anti-pro Caspase 9 antibody [E84] (ab32068)
Predicted band size : 46 kDa
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 recognize CASP9 when CASP9 knockout samples were used, along with additional cross-reactive bands. 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 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.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-Pro-Caspase-9 RabMAb (ab32068).
Western blot - pro Caspase 9 antibody [E84] (ab32068)
Anti-pro 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 for Anti-pro Caspase 9 antibody [E84] (ab32068)
This product has been referenced in:
Wang C et al. JD enhances the anti-tumour effects of low-dose paclitaxel on gastric cancer MKN45 cells both in vitro and in vivo. Cancer Chemother Pharmacol78:971-982 (2016).
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