Recombinant Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-274538-anti-active-caspase-3-antibody-e83-77-western-blot.jpg "Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E83-77] to Cleaved Caspase-3
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Cleaved Caspase-3 antibody [E83-77]
See all Cleaved Caspase-3 primary antibodies -
Description
Rabbit monoclonal [E83-77] to Cleaved Caspase-3 -
Host species
Rabbit -
Specificity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information This antibody only detects the active (cleaved) form of Caspase-3 and does not recognize the pro form of Caspase-3. PubMedID 19789217 describes the detection of human cells injected into mice. The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway. -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-P or IP -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Synthetic peptide within Human Cleaved Caspase-3 aa 1-100 (N terminal). The exact sequence is proprietary. A synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).
Database link: P42574 -
Positive control
- WB: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate; Jurkat cell lysate (camptothecin treated); HeLa cell lysate (staurosporine treated). ICC/IF: Hela cells (staurosporine treated); Human Vascular endothelial cells.
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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. 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
E83-77 -
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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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32042 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 | (7) |
1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).
The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway. |
| ICC/IF | (5) |
1/100 - 1/250.
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| Notes |
|---|
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WB
1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa). The pro-caspase-3 is cleaved only when apoptosis event occurs. So, in order to detect active Caspase-3, we strongly suggest to induce your samples into apoptotic pathway. |
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ICC/IF
1/100 - 1/250. |
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. Triggers cell adhesion in sympathetic neurons through RET cleavage. -
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
- Omim: 600636 Human
- SwissProt: P42574 Human
- Unigene: 141125 Human
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Alternative names
- active caspase 3 antibody
- Apopain antibody
- CASP 3 antibody
see all
Images
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Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Lane 1: Wild type HAP1 + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 2: Wild type HAP1 + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lane 3: HAP1 CASP3 KO + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 4: HAP1 CASP3 KO + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lane 5: HeLa + DMSO for 24 hours, whole cell lysate (20 µg)
Lane 6: HeLa + 2uM Staurosporine (ab146588) for 24 hours, whole cell lysate (20 µg)
Lanes 1 - 6: Merged signal (red and green). Green - ab32042 observed at 17 kDa. Red - loading control, ab130007, observed at 130 kDa.ab32042 was shown to specifically react with CASP3 (Caspase-3) when CASP3 (Caspase-3) knockout samples were used. HAP1 wild-type and CASP3 (Caspase-3) knockout samples were subjected to SDS-PAGE. Ab32042 and ab130007 (Mouse anti vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
Cells were grown to confluency prior to treatment.
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![Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-320368-anti-active-caspase-3-antibody-e83-77-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)Image from Zhu ZX et al., PLoS One. 2015;10(12):e0144495. Fig 7.; doi: 10.1371/journal.pone.0144495. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/High-glucose induces apoptosis in human Vascular endothelial cells (VECs).
Apoptotic responses in VEC were analyzed by detection of cleaved-Caspase-3 immunofluorescence using ab32042. Cells were treated with low glucose (LG) or high glucose (HG) for 72 hours before treated with 100 ng/mL bFGF, 1 μM sp600125 (sp), or 1 μM U0126 (U) or 10 μM MnTmPyP for 1 hour. Bar = 100 μm.
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![Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-43718-anti-active-caspase-3-antibody-e83-77-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)Ab32042, at dilution of 1/100, staining HeLa (human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.
Left image: control.
Right image: staurosporine treated. -
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-251779-anti-active-caspase-3-antibody-e83-77-western-blot.jpg)
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)All lanes : Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1 : HeLa Whole Cell Lysate (2 uM Staurosporine, 4Hr) at 20 µg
Lane 2 : HeLa Whole Cell Lysate (untreated) at 20 µg
Lane 3 : Cleaved Caspase 3 (recombinant protein) at 0.1 µg
Secondary
All lanes : 800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 32 kDa
Additional bands at: 17 kDa (possible mature (processed) protein) -
![Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-43719-anti-active-caspase-3-antibody-e83-77-western-blot.jpg)
Western blot - Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)Lane 1 : anti Pro Caspase 3 at 1/10000 dilution
Lane 2 : anti Pro Caspase 3 at 1/10000 dilution
Lanes 3-4 : Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lanes 2 & 4 : Jurkat cell lysate + Camptothecin
Lane 3 : Jurkat cell lysate
Predicted band size: 32 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands. -
![Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)](https://www.abcam.com/ps/products/32/ab32042/Images/ab32042-9-benefits-of-recombinant-antibodies.png)
Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042)
Datasheets and documents
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SDS download
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Datasheet download
References (293)
ab32042 has been referenced in 293 publications.
- Tian Y et al. TRPM2-AS Promotes Bladder Cancer by Targeting miR-22-3p and Regulating GINS2 mRNA Expression. Onco Targets Ther 14:1219-1237 (2021). PubMed: 33658791
- Mao J et al. ASPP2 expression predicts the prognosis of patients with hepatocellular carcinoma after transcatheter arterial chemoembolization. Exp Ther Med 21:397 (2021). PubMed: 33680119
- Zha W et al. Role of the long non-coding RNA HOTAIR/miR-126 axis in an in vitro psoriasis model. Exp Ther Med 21:450 (2021). PubMed: 33747185
- Chu H et al. SERPINE2 feedback regulates EGF/EGFR signaling in human papillary thyroid carcinoma cells. Mol Med Rep 23:N/A (2021). PubMed: 33760135
- Sun S et al. Roles of the microRNA-338-3p/NOVA1 axis in retinoblastoma. Mol Med Rep 23:N/A (2021). PubMed: 33760207