Recombinant Anti-Caspase-3 antibody [EPR18297] (ab184787)
![Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-260591-anti-caspase-3-antibody-epr18297-western-blot.jpg "Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18297] to Caspase-3
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Caspase-3 antibody [EPR18297]
See all Caspase-3 primary antibodies -
Description
Rabbit monoclonal [EPR18297] to Caspase-3 -
Host species
Rabbit -
Specificity
ab184787 recognizes pro-Caspase 3 and potentially cross reacts with active caspases after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells -
Tested applications
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat whole cell lysate and Jurkat whole cell lysate treated with 1uM staurosporine for 4 hours. IHC-P: Human tonsil and cervical cancer tissues. IP: HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18297 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab184787 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 | (2) |
1/2000. Detects a band of approximately 32, 17 kDa (predicted molecular weight: 32 kDa).
The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17). |
| IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| IP |
1/80.
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| Notes |
|---|
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WB
1/2000. Detects a band of approximately 32, 17 kDa (predicted molecular weight: 32 kDa). The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17). |
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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IP
1/80. |
Target
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Function
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. -
Tissue specificity
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system. -
Sequence similarities
Belongs to the peptidase C14A family. -
Post-translational
modificationsCleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 836 Human
- Entrez Gene: 12367 Mouse
- Entrez Gene: 25402 Rat
- Omim: 600636 Human
- SwissProt: P42574 Human
- SwissProt: P70677 Mouse
- SwissProt: P55213 Rat
- Unigene: 141125 Human
see all -
Alternative names
- A830040C14Rik antibody
- Apopain antibody
- CASP 3 antibody
see all
Images
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Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Lane 1: Wild-type HAP1 cell lysate + Staurosporine (1μM for 4h)
Lane 2: Wild-type HAP1 cell lysate
Lane 3: Caspase-3 knockout HAP1 cell lysate + Staurosporine (1μM for 4h)
Lane 4: Caspase-3 knockout HAP1 cell lysate
Lanes 1 - 4: Merged signal (red and green). Green - ab184787 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab184787 was shown to recognise pro Caspase 3 when Caspase 3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase 3 knockout samples (± Staurosporine treatment) were subjected to SDS-PAGE. ab184787 and ab8245 (loading control to GAPDH) were diluted to 1/2000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
![Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-311842-anti-caspase-3-antibody-epr18297-western-blot.jpg)
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 0.7 µg/ml
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 1µM Staurosporine 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 1/100000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and diluting buffer: 5% NFDM/TBST.
ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples. -
![Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-243795-anti-caspase-3-antibody-epr18297-western-blot.jpg)
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution
Lane 1 : Untreated Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lane 2 : Jurkat whole cell lysates treated with 1uM staurosporine for 4 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 17,32 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
Specificity: interacts with full length pro-Caspase 3 and the p17 subunit.
The Caspase-3 precursor is first cleaved between D175 and S176 to produce the p11 subunit and p20 fragment. Subsequently, the p20 fragment is cleaved between D28 and S29 to generate the p17 subunit (Proc. Natl. Acad. Sci. USA. 93, 7464-7469 - PMID:8755496).
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![Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-275079-anti-caspase-3-antibody-epr18297-western-blot.jpg)
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)All lanes : Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse spleen tissue lysate
Lane 3 : Rat spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 32 kDaLanes 1 - 4: Merged signal (red and green). Green - ab184787 observed at 32 kDa. Red - loading control, ab18058, observed at 130 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab184787 and ab18058 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-243793-anti-caspase-3-antibody-epr18297-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on lymphocytes of tonsil 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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-243790-anti-caspase-3-antibody-epr18297-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on tumor cells of Human cervix cancer 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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-243788-anti-caspase-3-antibody-epr18297-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)active and pro Caspase 3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 1uM staurosporine for 4 hours with ab184787 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab184787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.
Lane 1: HeLa whole cell lysate treated with 1uM staurosporine for 4 hours 10 µg (Input).
Lane 2: ab184787 IP in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184787 in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 3 minutes.
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![Anti-Caspase-3 antibody [EPR18297] (ab184787)](https://www.abcam.com/ps/products/184/ab184787/Images/ab184787-8-benefits-of-recombinant-antibodies.png)
Anti-Caspase-3 antibody [EPR18297] (ab184787)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (76)
ab184787 has been referenced in 76 publications.
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- Nie Z et al. The protective effects of resveratrol pretreatment in cyclophosphamide-induced rat ovarian injury: an vivo study. Gynecol Endocrinol N/A:1-6 (2021). PubMed: 33594937
- Xu XS et al. SCARF1 promotes M2 polarization of Kupffer cells via calcium-dependent PI3K-AKT-STAT3 signalling to improve liver transplantation. Cell Prolif 54:e13022 (2021). PubMed: 33686740