Recombinant Anti-Caspase-3 p12 antibody [EPR16888] (ab179517)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16888] to Caspase-3 p12
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Caspase-3 p12 antibody [EPR16888]
See all Caspase-3 p12 primary antibodies -
Description
Rabbit monoclonal [EPR16888] to Caspase-3 p12 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more 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: HeLa, PC12 and NIH/3T3 whole cell lysates. Mouse and Rat brain, hippocampus,spinal cord, cerebellum, cerebral cortex, liver and heart tissue lysates, human brain, liver, hypothalamus, heart and cerebellum tissue lysates. IHC-P: Human tonsil, mouse testis and rat kidney tissues. ICC/IF: K562 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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16888 -
Isotype
IgG
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab179517 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 |
1/1000. Detects a band of approximately 32 ,29, 12 kDa (predicted molecular weight: 32, 29, 12 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/150.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000. Detects a band of approximately 32 ,29, 12 kDa (predicted molecular weight: 32, 29, 12 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/150. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
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
- SwissProt: P42574 Human
- SwissProt: P70677 Mouse
- SwissProt: P55213 Rat
- Unigene: 141125 Human
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Form
Caspase-3 p12 is from aa 176-277 of P42574. -
Alternative names
- Apopain antibody
- CASP-3 antibody
- CASP3 antibody
see all
Images
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : Mouse Alzheimer's disease brain tissue lysate
Lane 2 : Mouse brain cancer tissue lysate
Lane 3 : Mouse hippocampus tissue lysate
Lane 4 : Mouse spinal cord tissue lysate
Lane 5 : Mouse cerebellum tissue lysate
Lane 6 : Mouse cerebral cortex tissue lysate
Lane 7 : Mouse hypothalamus tissue lysate
Lane 8 : Mouse heart tissue lysate
Lane 9 : Mouse liver tissue lysate
Lane 10 : Human brain tissue lysate
Lane 11 : Human liver tissue lysate
Lane 12 : Human hypothalamus tissue lysate
Lane 13 : Human heart tissue lysate
Lane 14 : Human cerebellum tissue
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 32, 29, 12 kDa
Observed band size: 27-32 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as loading control.
Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat hippocampus tissue lysate
Lane 3 : Rat spinal cord tissue lysate
Lane 4 : Rat cerebellum tissue lysate
Lane 5 : Rat cerebral cortex tissue
Lane 6 : Rat hypothalamus tissue
Lane 7 : Rat heart tissue lysate
Lane 8 : Rat liver tissue lysate
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 32, 29, 12 kDa
Observed band size: 27-32 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as loading control.
Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates treated with staurosporine 1uM for 4hr
Lane 2 : HeLa untreated whole cell lysates
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, 29, 12 kDa
Observed band size: 12,29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.
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Intracellular Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) with purified ab179517 at 1/240 dilution (Red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Caspase-3 p12 with ab179517 at 1/150 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1: ab179517 at 1/150 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) treated with staurosporine 1uM for 4h
Lane 2 : Untreated NIH/3T3 whole cell lysates
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, 29, 12 kDa
Observed band size: 12,29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3; 12-kDa band is Caspase-3 subunit p12.
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Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/5000 dilution + Mouse spleen lysates at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 32, 29, 12 kDa
Observed band size: 29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3.
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All lanes : Anti-Caspase-3 p12 antibody [EPR16888] (ab179517) at 1/1000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Mouse heart lysates
Lane 3 : Rat brain lysates
Lane 4 : Rat heart lysates
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, 29, 12 kDa
Observed band size: 29,32 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
32 and 29 kDa bands represent procaspase-3.
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining in germinal center, weaker staining in the mantle zone (MZ) lymphocytes is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (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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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on leydig cells and spermatogonial cells is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (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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Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Caspase-3 p12 with ab179517 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Granular staining on epithelium from kidney is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution.
[1] Afshin Samali, Boris Zhivotovsky, Dean P. Jonesa, Sten Orreniusa. Detection of pro-caspase-3 in cytosol and mitochondria of various tissues (rat). FEBS Letters 431 (1998) 167-169.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (23)
ab179517 has been referenced in 23 publications.
- Amadeu SO et al. Arthrospira (Spirulina) platensis feeding reduces the early stage of chemically induced rat colon carcinogenesis. Br J Nutr 129:395-405 (2023). PubMed: 35506448
- Xie ZQ et al. Capsaicin suppresses hepatocarcinogenesis by inhibiting the stemness of hepatic progenitor cells via SIRT1/SOX2 signaling pathway. Cancer Med 11:4283-4296 (2022). PubMed: 35674129
- Yang H et al. CRISPR/Cas9‑induced saturated mutagenesis identifies Rad51 haplotype as a marker of PARP inhibitor sensitivity in breast cancer. Mol Med Rep 26:N/A (2022). PubMed: 35713220
- Huo X et al. Lentinan Enhances the Function of Oxaliplatin on the Esophageal Tumors by Persuading Immunogenic Cell Death. Comput Math Methods Med 2022:2296574 (2022). PubMed: 35844448
- Zhu S et al. Injectable conductive gelatin methacrylate / oxidized dextran hydrogel encapsulating umbilical cord mesenchymal stem cells for myocardial infarction treatment. Bioact Mater 13:119-134 (2022). PubMed: 35224296