Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Caspase-9 antibody [EPR18108] (ab185719)

Submit a review Submit a question References (5)
Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Immunoprecipitation - Anti-Caspase-9 antibody [EPR18108] (ab185719)
  • Anti-Caspase-9 antibody [EPR18108] (ab185719)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18108] to Caspase-9
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Caspase-9 antibody [EPR18108]
    See all Caspase-9 primary antibodies

  • Description

    Rabbit monoclonal [EPR18108] to Caspase-9

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment within Human Caspase-9 aa 100-350. The exact sequence is proprietary.
    Database link: P55211

  • Positive control

    • WB: HeLa whole cell lysates untreated and treated with staurosporine 1uM for 4 hours; NIH/3T3 whole cell lysates untreated and treated with staurosporine1uM for 4 hours; Human fetal brain, fetal heart, fetal kidney and fetal liver and pancreas lysates; Mouse heart and kidney lysates. IP: HeLa whole cell lysate treated with staurosporine 1uM for 4 hours.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab185719 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 1/1000. Detects a band of approximately 46, 37, 35 kDa (predicted molecular weight: 46 kDa).
IP 1/40.

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 [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Caspase-9 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Jurkat cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab185719 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab185719 was shown to specifically react with Caspase-9 when Caspase-9 knockout samples were used. Wild-type and Caspase-9 knockout samples were subjected to SDS-PAGE. ab185719 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/10000 dilution

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 46 kDa
      Observed band size: 35,37,46 kDa
      why is the actual band size different from the predicted?


      Exposure time: 10 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/2500 dilution

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 46 kDa
      Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?


      Exposure time: 30 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution

      Lane 1 : Human fetal brain lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/5000 dilution

      Lane 1 : Human pancreas lysate
      Lane 2 : Human fetal liver lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


      Exposure time: 30 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Western blot - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      All lanes : Anti-Caspase-9 antibody [EPR18108] (ab185719) at 1/1000 dilution

      Lane 1 : Mouse heart lysate
      Lane 2 : Mouse kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 46 kDa
      Observed band size: 37,39,49 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Immunoprecipitation - Anti-Caspase-9 antibody [EPR18108] (ab185719)

      Caspase-9 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with staurosporine 1uM for 4 hours with ab185719 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab185719 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

      Lane 1: HeLa whole cell lysate treated with staurosporine 10 µg (Input).

      Lane 2: ab185719 IP in HeLa whole cell lysate treated with staurosporine.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185719 in HeLa whole cell lysate treated with staurosporine.
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    • Anti-Caspase-9 antibody [EPR18108] (ab185719)
      Anti-Caspase-9 antibody [EPR18108] (ab185719)

    References (5)

    Publishing research using ab185719? Please let us know so that we can cite the reference in this datasheet.

    ab185719 has been referenced in 5 publications.

    • Huang Y  et al. Dehydrocorydaline inhibits the tumorigenesis of breast cancer MDA-MB-231 cells. Mol Med Rep 22:43-50 (2020). PubMed: 32377708
    • Yagensky O  et al. Increased expression of heme-binding protein 1 early in Alzheimer's disease is linked to neurotoxicity. Elife 8:N/A (2019). PubMed: 31453805
    • Wang X  et al. Silence of lncRNA ANRIL represses cell growth and promotes apoptosis in retinoblastoma cells through regulating miR-99a and c-Myc. Artif Cells Nanomed Biotechnol 47:2265-2273 (2019). PubMed: 31184221
    • Zhong X  et al. Downregulation of microRNA-34a inhibits oxidized low-density lipoprotein-induced apoptosis and oxidative stress in human umbilical vein endothelial cells. Int J Mol Med 42:1134-1144 (2018). PubMed: 29750293
    • Luff SA  et al. Role of p53 and transcription-independent p53-induced apoptosis in shear-stimulated megakaryocytic maturation, particle generation, and platelet biogenesis. PLoS One 13:e0203991 (2018). PubMed: 30231080

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    There are currently no Customer reviews or Questions for ab185719.
    Please use the links above to contact us or submit feedback about this product.

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