Anti-Caspase-9 antibody [EPR18107] (ab202068) is a rabbit monoclonal antibody that is used to detect Caspase-9 in Western Blot, IP, IHC-P, ICC/IF. Suitable for Human, Mouse samples.
- Specificity confirmed with Caspase-9 knockout cell line validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
IP | WB | ICC/IF | IHC-P | |
---|---|---|---|---|
Human | Tested | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/80 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/2000 | Notes - |
Species Human | Dilution info 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/300 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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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.
MCH6, CASP9, Caspase-9, CASP-9, Apoptotic protease Mch-6, Apoptotic protease-activating factor 3, ICE-like apoptotic protease 6, APAF-3, ICE-LAP6
Anti-Caspase-9 antibody [EPR18107] (ab202068) is a rabbit monoclonal antibody that is used to detect Caspase-9 in Western Blot, IP, IHC-P, ICC/IF. Suitable for Human, Mouse samples.
- Specificity confirmed with Caspase-9 knockout cell line validation
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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False colour image of Western blot: Anti-Caspase-9 antibody [EPR18107] staining at 1/2000 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, ab202068 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 [EPR18107] (ab202068) at 1/2000 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
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)
Predicted band size: 46 kDa
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) (Goat Anti-Rabbit IgG H&L (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 Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-CDT1/DUP antibody [EPR17891] ab202067 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
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) (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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Green - target observed at 46 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
This western blot image is a comparison between ab202068 and a competitor's top cited rabbit polyclonal antibody.
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068)
Predicted band size: 46 kDa
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.
All lanes: Immunoprecipitation - Anti-Caspase-9 antibody [EPR18107] (ab202068)
Predicted band size: 46 kDa
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/10000 dilution
Lane 1: Untreated C2C12 (Mouse myoblast cell line) whole cell lysate at 10 µg
Lane 2: C2C12 (Mouse myoblast cell line) treated with staurosporine 1uM for 4 hours whole cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 37 kDa, 39 kDa, 46 kDa
Exposure time: 3min
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/2000 dilution
Lane 1: Human fetal brain lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 3min
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/2000 dilution
Lane 1: Human fetal kidney lysate at 10 µg
Lane 2: Human fetal liver lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 46 kDa
Exposure time: 30s
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Caspase-9 antibody [EPR18107] (ab202068) at 1/50000 dilution
Lane 1: Untreated HeLa (human epithelial cells from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 2: HeLa (human epithelial cells from cervix adenocarcinoma) treated with staurosporine 1 µM for 4 hours whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 46 kDa
Observed band size: 35 kDa, 37 kDa, 46 kDa
Image collected and cropped by CiteAb under a CC-BY license from the publication
Caspase-9 western blot using anti-Caspase-9 antibody [EPR18107] ab202068. Publication image and figure legend from Mesmar, F., Dai, B., et al., 2019, Cancer Med, PubMed 31568691.
ab202068 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab202068 please see the product overview.
GPER1 mediates anti‐proliferative effects in pancreatic cancer. A, GPER1 agonist (G1, 2 μmol/L) reduces cell proliferation of both cell lines (MTT assay, 72 h), which is attenuated upon addition of GPER1 antagonist (G15). B, G1 induces PARP, caspase 3, and caspase 9 cleavage in both MiaPaCa2 and PANC1. C, Propidium iodide (PI) and Annexin V staining demonstrate apoptosis in PANC1 cells after G1 treatment (2 μmol/L, 48 h). Error bar represents SEM and unpaired two‐tailed t test was used to test significance (*P < .05, **P < .01, ***P < .001)
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