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Rabbit Recombinant Monoclonal Caspase-9 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 248 publications.


Images

Western blot - Anti-Caspase-9 antibody [E23] (AB32539), expandable thumbnail
  • Western blot - Anti-Caspase-9 antibody [E23] (AB32539), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (AB32539), expandable thumbnail
  • Immunoprecipitation - Anti-Caspase-9 antibody [E23] (AB32539), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] (AB32539), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species
Human
Dilution info
1/50 - 1/250
Notes

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

Tested
Tested

Species
Human
Dilution info
1/80 - 1/100
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000 - 1/10000
Notes

We recommend overnight incubation at 4°C.

Tested
Tested

Species
Human
Dilution info
1/500
Notes

For unpurified use at 1/50.

Tested
Tested

Species
Human
Dilution info
1/100 - 1/250
Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

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Target data

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 effector caspases caspase-3 (CASP3) or caspase-7 (CASP7). Promotes DNA damage-induced apoptosis in a ABL1/c-Abl-dependent manner. Proteolytically cleaves poly(ADP-ribose) polymerase (PARP). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758105, PubMed:36758106). Isoform 2. Lacks activity is an dominant-negative inhibitor of caspase-9.

Alternative names

MCH6, CASP9, Caspase-9, CASP-9, Apoptotic protease Mch-6, Apoptotic protease-activating factor 3, ICE-like apoptotic protease 6, APAF-3, ICE-LAP6

Recommended products

Rabbit Recombinant Monoclonal Caspase-9 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 248 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
E23
Purification technique
Affinity purification Protein A
Specificity

This antibody should recognise both the pro-[40kDa] form and p35 cleaved form of Caspase-9.

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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

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 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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

  • Western blot - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Western blot - Anti-Caspase-9 antibody [E23] (ab32539)

    False colour image of Western blot: Anti-Caspase-9 antibody [E23] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32539 was shown to bind specifically to Caspase-9. A band was observed at 45 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CASP9 knockout cell line Human CASP9 knockout THP-1 cell line ab276122 (knockout cell lysate ab284219). To generate this image, wild-type and CASP9 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-Caspase-9 antibody [E23] (ab32539) at 1/1000 dilution

    Lane 1: Wild-type THP-1 cell lysate at 20 µg

    Lane 2: CASP9 knockout THP-1 cell lysate at 20 µg

    Lane 2: Western blot - Human CASP9 knockout THP-1 cell line (Human CASP9 knockout THP-1 cell line ab276122)

    Lane 3: HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 46 kDa

    Observed band size: 45 kDa

  • Western blot - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Western blot - Anti-Caspase-9 antibody [E23] (ab32539)

    Lanes:
    Lane 1: Wild-type HAP1 whole cell lysate (40 μg)
    Lane 2: CASP9 knockout HAP1 whole cell lysate (40 μg)
    Lane 3: HeLa whole cell lysate (40 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32539 observed at 45 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab32539 was shown to specifically recognize Caspase-9 in wild type HAP1 cells along with additional cross-reactive bands. No band was observed when Caspase-9 knockout samples were examined. Wild-type and Caspase-9 knockout samples were subjected to SDS-PAGE. ab32539 and Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Caspase-9 antibody [E23] (ab32539)

    Predicted band size: 46 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539)

    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. Goat Anti-Rabbit IgG H&L (HRP) 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.

  • Immunoprecipitation - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Immunoprecipitation - Anti-Caspase-9 antibody [E23] (ab32539)

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.

    All lanes: Immunoprecipitation - Anti-Caspase-9 antibody [E23] (ab32539)

    Predicted band size: 46 kDa

    Observed band size: 46 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Caspase-9 antibody [E23] (ab32539)

    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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/500) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow Cytometry (Intracellular) - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Caspase-9 antibody [E23] (ab32539)

    Intracellular 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.

  • Western blot - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Western blot - Anti-Caspase-9 antibody [E23] (ab32539)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Caspase-9 antibody [E23] (ab32539) at 1/1000 dilution

    Lane 1: HeLa whole cell lysate - treated with Camptothecin at 10 µg

    Lane 2: HeLa whole cell lysate - untreated at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 46 kDa

    Observed band size: 35 kDa, 46 kDa

  • Flow Cytometry (Intracellular) - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Caspase-9 antibody [E23] (ab32539)

    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 (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.

  • Western blot - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Western blot - Anti-Caspase-9 antibody [E23] (ab32539)

    All lanes: Western blot - Anti-Caspase-9 antibody [E23] (ab32539) at 1/20000 dilution

    Lane 1: Jurkat cell lysate - untreated

    Lane 2: Jurkat cell lysate - treated with Camptothecin

    Predicted band size: 46 kDa

    Observed band size: 28 kDa, 48 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-9 antibody [E23] (ab32539)

    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.

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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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