Anti-Caspase-3 antibody [31A1067] (ab13585)
Key features and details
- Mouse monoclonal [31A1067] to Caspase-3
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
Overview
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Product name
Anti-Caspase-3 antibody [31A1067]
See all Caspase-3 primary antibodies -
Description
Mouse monoclonal [31A1067] to Caspase-3 -
Host species
Mouse -
Specificity
ab13585 recognizes an active form of Caspase 3 after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein corresponding to Human Caspase-3 aa 1-277.
Database link: P42574 -
Positive control
- Staurosporine-treated HeLa or Jurkat cell lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.4
Preservative: 0.05% Sodium azide
Constituents: 99% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
31A1067 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab13585 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 |
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WB |
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 31 kDa.
The antibody detects both pro Caspase 3 (~32 kDa) and the large subunit of the active/cleaved form (~14-21 kDa) of Caspase 3. The large subunit of the cleaved form may appear as one or two or even as a stack of bands depending on the presence or absence of the Caspase 3 pro-domain. |
Notes |
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WB
Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 31 kDa. The antibody detects both pro Caspase 3 (~32 kDa) and the large subunit of the active/cleaved form (~14-21 kDa) of Caspase 3. The large subunit of the cleaved form may appear as one or two or even as a stack of bands depending on the presence or absence of the Caspase 3 pro-domain. |
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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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 - ab13585 observed at 32 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab13585 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. ab13585 at a concentration of 1 µg/ml and ab8245 (loading control to GAPDH) diluted to 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
Western blot analysis of Caspase 3 in HeLa cells using ab13585. Cells were treated with 2 uM staurosporine for different time periods. Caspase 3 activation is detected in Western blots by the presence of Caspase 3 cleavage fragments.
ab13585 detected both pro (full-length) and active (cleaved) Caspase 3, depending on the treatment time points. Pro Caspase 3 is detected at ~32 kDa. Active/cleaved Caspase 3 (large subunit) is detected at ~14-21 kDa as one or more bands. -
All lanes : Anti-Caspase-3 antibody [31A1067] (ab13585) at 5 µg/ml
Lane 1 : Human brain lysate
Lanes 2 & 12 : Human heart lysate
Lane 3 : Human intestine lysate
Lane 4 : Human kidney lysate
Lane 5 : Human liver lysate
Lane 6 : Human lung lysate
Lane 7 : Human muscle lysate
Lane 8 : Human stomach lysate
Lane 9 : Human spleen lysate
Lane 10 : Human ovary lysate
Lane 11 : Human testis lysate
Lane 13 : Mouse heart lysate
Lane 14 : Rat heart lysate
Predicted band size: 31 kDa
Lanes 12, 13 and 14 demonstrate the species cross-reactivity of the antibody in Human, mouse and rat heart lysate, respectively. -
All lanes : Anti-MDC1 antibody (ab13858) at 1 µg/ml
Lane 1 : Hela whole cell lysate (Staurosporine treated, 2uM/4hr)
Lane 2 : Hela whole cell lysate (untreated control)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 31 kDa
Additional bands at: 17 kDa (possible mature (processed) protein), 19 kDa (possible mature (processed) protein), 32 kDa (possible immature (unprocessed))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 ab13585 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Datasheets and documents
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SDS download
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Datasheet download
References (101)
ab13585 has been referenced in 101 publications.
- Tang J et al. Circ_0006220 Contributes to NSCLC Progression through miR-342-3p/GOT2 Axis. Ann Thorac Cardiovasc Surg 29:11-22 (2023). PubMed: 36575008
- Lin L et al. Long non-coding RNA MEG3 promotes autophagy and apoptosis of nasopharyngeal carcinoma cells via PTEN up-regulation by binding to microRNA-21. J Cell Mol Med 25:61-72 (2021). PubMed: 33332708
- Liang R et al. Lycium barbarum polysaccharide protects ARPE‑19 cells against H2O2‑induced oxidative stress via the Nrf2/HO‑1 pathway. Mol Med Rep 24:N/A (2021). PubMed: 34490478
- Che F et al. LncRNA MALAT1 induced by hyperglycemia promotes microvascular endothelial cell apoptosis through activation of the miR-7641/TPR axis to exacerbate neurologic damage caused by cerebral small vessel disease. Ann Transl Med 9:1762 (2021). PubMed: 35071456
- Jin S et al. 5-Azacitidine Induces NOXA to Prime AML Cells for Venetoclax-Mediated Apoptosis. Clin Cancer Res 26:3371-3383 (2020). PubMed: 32054729