Overview

  • Product name

    Anti-Caspase-9 antibody [EPR18868]
    See all Caspase-9 primary antibodies
  • Description

    Rabbit monoclonal [EPR18868] to Caspase-9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Rat Caspase-9 aa 1-200. The exact sequence is proprietary.
    Database link: Q9JHK1

  • Positive control

    • WB: C6 and NIH/3T3 whole cell lysates; Mouse brain, heart and spleen lysates; Rat brain and heart lysates. IP: NIH/3T3 and C6 treated with 1µM staurosporine for 4h whole cell lysates.
  • General 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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab184786 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
IP 1/30.
WB 1/1000. Detects a band of approximately 50, 39, 37 kDa (predicted molecular weight: 50, 39, 37 kDa).

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

  • All lanes : Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

    Lane 1 : Untreated C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 0.5 hour
    Lane 3 : C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 1 hour
    Lane 4 : C6 whole cell lysate treated with 1 µm staurosporine for 4 hours
    Lane 5 : C6 whole cell lysate treated with 50 µm Z-VAD-FMK for 1 hour, then treated with 1 µM staurosporine for 4 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 50, 39, 37 kDa
    Observed band size: 37,39,50 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The p39 and p37 subunits were inhibited when the caspase activity was blocked by the caspase inhibitor, Z-VAD-FMK.

  • All lanes : Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embyro fibroblast cell line) whole cell lysate
    Lane 2 : NIH/3T3 whole cell lysate treated with 1 µM staurosporine for 4 hours

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 50, 39, 37 kDa
    Observed band size: 37,39,50 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Caspase-9 antibody [EPR18868] (ab184786) at 1/1000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 50, 39, 37 kDa
    Observed band size: 37,39,50 kDa why is the actual band size different from the predicted?


    Exposure time: 20 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Caspase 9 was immunoprecipitated from 1mg of NIH/3T3 whole cell lysate (Mouse embyro fibroblast cells) treated with 1μM staurosporine for 4h with ab184786 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184786 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h,10ug (Input).
    Lane 2: ab184786 IP in NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184786 in NIH/3T3 whole cell lysate treated with 1μM staurosporine for 4h.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 10 seconds.

     

    The product has better affinity to procaspase-9. The cleaved form p37 could not be observed even with 3 minutes exposure time.

  • Caspase 9 was immunoprecipitated from 1mg of C6 whole cell lysate (Rat glial tumor cells) treated with 1μM staurosporine for 4h with ab184786 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab184786 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: C6 whole cell lysate treated with 1μM staurosporine for 4h, 10ug (Input).
    Lane 2: ab184786 IP in C6 whole cell lysate treated with 1μM staurosporine for 4h.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184786 in C6 whole cell lysate treated with 1μM staurosporine for 4h.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

     

    The product has better affinity to procaspase-9. The cleaved form p37 could be observed with longer exposure time.

References

This product has been referenced in:

  • Yen JH  et al. Activation of dynamin-related protein 1 - dependent mitochondria fragmentation and suppression of osteosarcoma by cryptotanshinone. J Exp Clin Cancer Res 38:42 (2019). Read more (PubMed: 30691497) »
  • E L  et al. MicroRNA-144 attenuates cardiac ischemia/reperfusion injury by targeting FOXO1. Exp Ther Med 17:2152-2160 (2019). Read more (PubMed: 30783480) »
See all 10 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Rat Cell lysate - whole cell (PC12 cell)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
PC12 cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C

Miss. Ji Yeon Lee

Verified customer

Submitted Jan 25 2019

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