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  1. Link

    caspase-9-antibody-e84-ab32068.pdf

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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 [E84] (ab32068)

  • Datasheet
Submit a review Submit a question References (14)

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Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
  • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)
  • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
  • Anti-Caspase-9 antibody [E84] (ab32068)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E84] to Caspase-9
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-Caspase-9 antibody [E84]
    See all Caspase-9 primary antibodies
  • Description

    Rabbit monoclonal [E84] to Caspase-9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Caspase-9 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • Jurkat cell lysate
  • 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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E84
  • 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
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-Caspase-9 antibody [E84] - BSA and Azide free (ab247220)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

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

    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32068 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
1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).
IHC-P
1/25. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/2000. Detects a band of approximately 46 kDa (predicted molecular weight: 46 kDa).
IHC-P
1/25. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC or IP.

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

    • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
      Western blot - Anti-Caspase-9 antibody [E84] (ab32068)

      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 specifically recognize CASP9 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CASP9 knockout samples were examined. 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 1/500 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
      Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
      All lanes : Anti-Caspase-9 antibody [E84] (ab32068) at 1/1000 dilution

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

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 46 kDa
      Additional bands at: 35 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 3 minutes


      Blocking/dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E84] (ab32068)

      Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-Pro-Caspase-9 RabMAb (ab32068).

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
      Western blot - Anti-Caspase-9 antibody [E84] (ab32068)
      Anti-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

    • Anti-Caspase-9 antibody [E84] (ab32068)
      Anti-Caspase-9 antibody [E84] (ab32068)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (14)

    Publishing research using ab32068? Please let us know so that we can cite the reference in this datasheet.

    ab32068 has been referenced in 14 publications.

    • Zhao J  et al. Salidroside mitigates hydrogen peroxide-induced injury by enhancement of microRNA-27a in human trabecular meshwork cells. Artif Cells Nanomed Biotechnol 47:1758-1765 (2019). PubMed: 31062616
    • Meng FC & Lin JK Liquiritigenin Inhibits Colorectal Cancer Proliferation, Invasion, and Epithelial-to-Mesenchymal Transition by Decreasing Expression of Runt-Related Transcription Factor 2. Oncol Res 27:139-146 (2019). PubMed: 29471888
    • Yang Q  et al. Lidocaine alleviates cytotoxicity-resistance in lung cancer A549/DDP cells via down-regulation of miR-21. Mol Cell Biochem 456:63-72 (2019). PubMed: 30644017
    • Sun T  et al. Knockdown of Long Non-Coding RNA RP11-445H22.4 Alleviates LPS-Induced Injuries by Regulation of MiR-301a in Osteoarthritis. Cell Physiol Biochem 45:832-843 (2018). PubMed: 29414810
    • Gu LL  et al. Oxymatrine Causes Hepatotoxicity by Promoting the Phosphorylation of JNK and Induction of Endoplasmic Reticulum Stress Mediated by ROS in LO2 Cells. Mol Cells 41:401-412 (2018). PubMed: 29754474
    View all Publications for this product

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