Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Caspase-9 antibody [EPR18868] (ab184786)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18868] to Caspase-9
  • Suitable for: IP, WB
  • Reacts with: Mouse, Rat

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 & Species

    Application Species
    IP
    Mouse
    Rat
    WB
    Mouse
    Rat
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

     

     

    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-Caspase-9 antibody [EPR18868] (ab184786)
    Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    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.

  • Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    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.

  • Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    Western blot - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    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.

  • Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)

    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.

  • Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)
    Immunoprecipitation - Anti-Caspase-9 antibody [EPR18868] (ab184786)

    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.

  • Anti-Caspase-9 antibody [EPR18868] (ab184786)
    Anti-Caspase-9 antibody [EPR18868] (ab184786)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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