Recombinant Anti-Cleaved Caspase-3 antibody [E83-77] - BSA and Azide free (ab208003)
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 - BSA and Azide free
- 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] - BSA and Azide free
See all Cleaved Caspase-3 primary antibodies -
Description
Rabbit monoclonal [E83-77] to Cleaved Caspase-3 - BSA and Azide free -
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.
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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. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat cell lysate, treated wild type HeLa cell lysate. ICC/IF: Hela cells.
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General notes
ab208003 is the carrier-free version of ab32042.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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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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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab208003 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 at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 32 kDa). |
ICC/IF
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
- 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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All lanes : Anti-Cleaved Caspase-3 antibody [E83-77] (ab32042) at 1/500 dilution
Lane 1 : Wild-type HeLa Treated Staurosporine (2uM, 4h) cell lysate
Lane 2 : CASP3 knockout HeLa Treated Staurosporine (2uM, 4h) cell lysate
Lane 3 : Wild-type HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate
Lane 4 : CASP3 knockout HeLa Vehicle Control Staurosporine (0uM, 4h) cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 16,28 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32042).
Anti-CASP3 antibody [E83-77] (ab32042) staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32042 was shown to bind specifically to CASP3. A band was observed at 16/28 kDa in treated wild-type HeLa cell lysates with no signal observed at this size in CASP3 knockout cell line. To generate this image, wild-type and CASP3 knockout HeLa 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Ab32042, at dilution of 1/100, staining HeLa cells by Immunofluorescence.
Left image: control.
Right image: staurosporine treated.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32042).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (4)
ab208003 has been referenced in 4 publications.
- Liu Y et al. Targeted delivery of irinotecan to colon cancer cells using epidermal growth factor receptor-conjugated liposomes. Biomed Eng Online 21:53 (2022). PubMed: 35918704
- Yang Y et al. dRTEL1 is essential for the maintenance of Drosophila male germline stem cells. PLoS Genet 17:e1009834 (2021). PubMed: 34644293
- Xu J et al. NEDD4 protects vascular endothelial cells against Angiotensin II-induced cell death via enhancement of XPO1-mediated nuclear export. Exp Cell Res 383:111505 (2019). PubMed: 31326389
- Shen J et al. Lycorine inhibits glioblastoma multiforme growth through EGFR suppression. J Exp Clin Cancer Res 37:157 (2018). PubMed: 30016965