Anti-active Caspase 3 antibody (ab13847)
- Product nameAnti-active Caspase 3 antibodySee all active Caspase 3 primary antibodies ...
- DescriptionRabbit polyclonal to active Caspase 3
- SpecificityAb13847 has been batch tested in WB using recombinant active Caspase 3 only. Not all batches may detect endogenous active caspase 3 in WB. However, some customers have successfully used ab13847 on endogenous lysates. Please contact Abcam Scientific Support for more information.
- Tested applicationsICC/IF, IHC-P, IHC-Fr, Flow Cyt, ICC, IHC (PFA fixed), IHC-FoFr, WB more details
- Species reactivityReacts with: Mouse, Rat, Human, Pig, Xenopus laevis, Fruit fly (Drosophila melanogaster), Indian Muntjac, Zebrafish, Rhesus monkey
Predicted to work with: Dog, Chinese Hamster
Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human active Caspase 3.
(Peptide available as ab13848.)
- Positive controlThis antibody gave a positive signal in WB with active Caspase 3 recombinant protein.
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab13847 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use a concentration of 5 µg/ml.|
|IHC-P||IHC-P: 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-Fr||IHC-Fr: Use at an assay dependent concentration.|
|Flow Cyt||Flow Cyt: 1/500.|
|ICC||ICC: Use at an assay dependent dilution.|
|IHC (PFA fixed)||IHC (PFA fixed): 1/300.|
|WB||WB: Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 17 kDa.Can be blocked with active Caspase 3 peptide (ab13848).|
- RelevanceThe caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10). The processed form of Caspase 3 consists of large (17kDa) and small (12kDa) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 Ser29 and Asp175 Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein. In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (eg mantle zone B cells). Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.
- Cellular localizationCytoplasmic
- Entrez Gene: 836 Human
- Entrez Gene: 12367 Mouse
- Entrez Gene: 397244 Pig
- Entrez Gene: 25402 Rat
- Entrez Gene: 140621 Zebrafish
- Omim: 600636 Human
- SwissProt: P42574 Human
- SwissProt: P70677 Mouse
- SwissProt: Q95ND5 Pig
- SwissProt: P55213 Rat
- SwissProt: P55866 Xenopus laevis
- Unigene: 141125 Human
- Unigene: 10562 Rat
- APOPAIN antibodyApopain precursor antibodyCASP3 antibody
- Caspase 3 apoptosis related cysteine protease antibodyCaspase3 antibodyCPP 32 antibodyCPP32B antibodyCysteine protease CPP32 antibodyHuman cysteine protease CPP32 isoform alpha mRNA complete cds antibodyICE3 antibodyLICE antibodyPARP cleavage protease antibodySCA1 antibodySREBP cleavage activity 1 antibodyYama antibodyYama protein antibody
Anti-active Caspase 3 antibody images
ab13847 was used in IHC of frozen sections from a rat brain with a kainite lesion. The non lesionned contralateral site serves as a negative control. The sections were fixed with paraformaldehyde. The tissue was perfused with 4% PFA and embedded in OCT compound and cut on the cryostat. The primary antibody was incubated for 12 hours at a dilution of 1/300. A biotin labelled secondary antibody was used at a dilution of 1/300.
ab13847 at a dilution of 1/500 staining Asynchronous HeLa cells by Immunocytochemistry. The antibody was incubated with the cells for 30 minutes and then detected using a Cy3 conjugated Goat Anti-Rabbit IgG (H+L). This antibody gives a predominantly nuclear focal staining pattern in all interphase nuclei investigated.
This image is courtesy of an Abreview by Kirk McManus submitted on 2 December 2005.
HeLa cells were fixed for 10 minutes at room temperature in 3.7% PFA and permeabilised in 0.1% Triton X-100/PBS then incubated with ab13847 (5µg/ml) for 1 hour at room temperature. The top panel shows control cells treated with DMSO. The bottom panel shows HeLa cells treated with 1 mM staurosporine for 4 hours to induce caspase-3 activation. ab13847 staining is shown in green and counterstaining with DAPI is shown in blue. 100x magnification.
The image shows the staining with ab13847 is very faint in the untreated control cultures, but very bright after activation of capsase-3 by treatment with the staurosporine. (N.B. in these cultures the nuclei are apoptotic).
ab13847 at 1/1000 staining Xenopus laevis wholemount and sectioned tails by IHC-P. The tissue was formaldehyde fixed and blocked prior to incubation with the antibody for 12 hours. An alkaline phosphatase conjugated goat anti-rabbit antibody was used as the secondary.
All lanes : Anti-active Caspase 3 antibody (ab13847) at 1 µg/ml
Lane 1 : Pro-Caspase 3 (inactive) recombinant
Lane 2 : Caspase 3 (active) recombinant
Lysates/proteins at 0.1 µg per lane.
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 17 kDa
Observed band size : 17 kDa
ICC/IF image of ab13847 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab13847, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293 and MCF7 cells at 5µg/ml.
ab13847 staining active caspase 3 in Human Jurkat cells by Flow Cytometry. Cells were prepared in a phosphate buffered solution containing 0.1% sodium azide with FBS fixed with paraformaldehyde and permeabilized with Triton X-100 and NP40. The sample was incubated with the primary antibody (1/100 in wash buffer) for 24 hours at 4°C. A FITC-conjugated Goat anti-rabbit Ig (1/100) was used as the secondary antibody.
Gating Strategy: Isolate cell population from plot of SSC-A / FSA-A
ab13847 staining active caspase 3 in HeLa cells treated with RTIL-13™ (ab120465), by ICC/IF. Increase in active caspase 3 expression correlates with increased concentration of RTIL-13™, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120465 (RTIL-13™) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab13847 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab13847 staining active caspase 3 in A549 cells treated with quercetin (ab120247), by ICC/IF. Increase in active caspase 3 expression correlates with increased concentration of quercetin, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120247 (quercetin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab13847 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab968999) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
References for Anti-active Caspase 3 antibody (ab13847)
This product has been referenced in:
- Gorgal T et al. Expression of apoptosis-regulating genes in the rat prostate following botulinum toxin type A injection. BMC Urol 12:1 (2012). IHC-FoFr ; Rat . Read more (PubMed: 22216975) »
- Pan D et al. Isolation and characterization of intestinal epithelial cells from normal and SIV-infected rhesus macaques. PLoS One 7:e30247 (2012). Read more (PubMed: 22291924) »