Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Caspase-9 antibody [EPR18107] - BSA and Azide free (ab222231)

Overview

  • Product name

    Anti-Caspase-9 antibody [EPR18107] - BSA and Azide free
    See all Caspase-9 primary antibodies
  • Description

    Rabbit monoclonal [EPR18107] to Caspase-9 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment aa 100-300. The exact sequence is proprietary.
    Database link: P55211

  • Positive control

    • WB: HeLa and C2C12 whole cell lysates; Human fetal brain, fetal heart, fetal kidney and fetal liver lysates. IHC-P: Human cervix carcinoma tissue. ICC/IF: HeLa cells. IP: HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.
  • General notes

    Ab222231 is the carrier-free version of ab202068. 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.

    ab222231 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab222231 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
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 46, 35, 37 kDa (predicted molecular weight: 46 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Caspase-9 with ab202068 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing cytoplasmic and nuclear staining on HeLa cell line. The expression increased after treatment with staurosporine (1uM) for 4 hours.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202067 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab202068).

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Caspase-9 with ab202068 at 1/300 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Cytoplasmic and nuclear staining on Human cervix carcinoma tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab202068).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Caspase-9 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) treated with staurosporine 1uM for 4 hours whole cell lysate with ab202068 at 1/80 dilution.

    Western blot was performed from the immunoprecipitate using ab202068 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 treated with staurosporine 1uM for 4 hours whole cell lysate10 µg (Input).

    Lane 2: ab202068 IP in HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202068 in HeLa treated with staurosporine 1uM for 4 hours whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab202068).

References

ab222231 has not yet been referenced specifically in any publications.

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