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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

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  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Immunoprecipitation - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
  • Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E23] to Caspase-9 - BSA and Azide free
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-Caspase-9 antibody [E23] - BSA and Azide free
    See all Caspase-9 primary antibodies
  • Description

    Rabbit monoclonal [E23] to Caspase-9 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-9 aa 250-350.
    Database link: P55211

  • Positive control

    • WB: Jurkat and HeLa cell lysates. IHC-P: Human skeletal muscle and cervical carcinoma tissues. ICC/IF: HepG2 cells. Flow Cyt: K562 cells. IP: HeLa whole cell lysate.
  • General notes

    ab219590 is the carrier-free version of ab32539 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab219590 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E23
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Caspases
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Cytochrome C
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Caspases
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Cytochrome C
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • PE Anti-Caspase-9 antibody [E23] (ab237431)
    • Anti-Caspase-9 antibody [E23] (ab32539)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant human Caspase-9 protein (Tagged) (ab198061)

Applications

Our Abpromise guarantee covers the use of ab219590 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 Use at an assay dependent concentration. Predicted molecular weight: 46 kDa.

We recommend overnight incubation at 4°C.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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.
  • Target information above from: UniProt accession P55211 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 842 Human
    • Omim: 602234 Human
    • SwissProt: P55211 Human
    • Unigene: 329502 Human
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue labelling Caspase-9 with purified ab32539 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

    • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Caspase-9 with purified ab32539 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

    • Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      Flow Cytometry analysis of K562 cells labelling Caspase-9 with purified ab32539 at 1/250 (red). Cells were fixed with 100% methanol. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

    • Immunoprecipitation - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Immunoprecipitation - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      ab32539 (purified) at 1/80 immunoprecipitating Caspase-9 in HeLa whole cell lysate.

      Lane 1 (input): HeLa whole cell lysate (10µg)

      Lane 2 (+): ab32539 + HeLa whole cell lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32539 in HeLa whole cell lysate.

      For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

    • Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Flow Cytometry - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      Overlay histogram showing K562 cells stained with unpurified ab32539 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab32539, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was anti-rabbit DyLight® 488 (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling Caspase 9 with unpurified ab32539 at a dilution of 1/50.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32539).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)
      Anti-Caspase-9 antibody [E23] - BSA and Azide free (ab219590)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (32)

    Publishing research using ab219590? Please let us know so that we can cite the reference in this datasheet.

    ab219590 has been referenced in 32 publications.

    • Shi C  et al. LncRNA OIP5-AS1 promotes cell proliferation and migration and induces angiogenesis via regulating miR-3163/VEGFA in hepatocellular carcinoma. Cancer Biol Ther 21:604-614 (2020). PubMed: 32329664
    • He J  et al. Epimedium Polysaccharide Ameliorates Benzene-Induced Aplastic Anemia in Mice. Evid Based Complement Alternat Med 2020:5637507 (2020). PubMed: 32256652
    • Sheng LX  et al. Design, Synthesis, and Evaluation of Novel 2-Methoxyestradiol Derivatives as Apoptotic Inducers Through an Intrinsic Apoptosis Pathway. Biomolecules 10:N/A (2020). PubMed: 31936880
    • Wang HW  et al. Reversal of multidrug resistance in leukemia cells using a transferrin-modified nanomicelle encapsulating both doxorubicin and psoralen. Aging (Albany NY) 12:6018-6029 (2020). PubMed: 32259795
    • Li S  et al. Circular RNA 0001785 regulates the pathogenesis of osteosarcoma as a ceRNA by sponging miR-1200 to upregulate HOXB2. Cell Cycle 18:1281-1291 (2019). PubMed: 31116090
    View all Publications for this product

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