|
|
|
|
Immunohistochemistry (Frozen sections) - active Caspase 3 antibody (ab13847)
(enlarge)
|
|
Immunocytochemistry/ Immunofluorescence - active Caspase 3 antibody (ab13847)
(enlarge)
|
|
Immunocytochemistry/ Immunofluorescence - active Caspase 3 antibody (ab13847)
(enlarge)
|
|
|
|
|
Product Name
|
active Caspase 3 antibody
|
| |
See all active Caspase 3 antibodies (4)...
|
|
Product type
|
Primary antibodies
|
|
Description
|
Rabbit polyclonal to active Caspase 3
|
|
Immunogen
|
Synthetic peptide conjugated to KLH derived from within residues 150 - 250 of Human active Caspase 3.
(Peptide available as ab13848.)
|
|
Reacts with
|
Hu, Ms, Rat, Dm, InMtj, RMk, Xl, Zfsh
|
|
Predicted to react
(species key)
|
ChHm, Dog, Pig
(Due to sequence homology)
|
|
Specificity
|
ab13847 is specific for the active form of Caspase 3. It detects both the large (17kDa) and small (12kDa) subunits from recombinant active Caspase 3. Ab13847 also detects the large subunit in Hela, A431 and Jurkat lysates.
|
|
Tested applications
(see key)
|
Flow Cyt, ICC/IF, IHC (PFA fixed), IHC-FoFr, IHC-P, WB
|
|
|
Abreviews
|
|
|
|
Application notes
(see key)
|
Recommended dilutions
Flow Cyt: 1/500.
ICC: Use at an assay dependent dilution.
ICC/IF: Use at a concentration of 5 µg/ml.
IHC-P: 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC (PFA fixed): 1/300.
IHC-FoFr: 1/300.
WB: Use at a concentration of 1 - 2 µg/ml. Predicted molecular weight: 17 kDa. Can be blocked with active Caspase 3 peptide (ab13848).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
|
|
Positive control
(see definition)
|
This antibody gave a positive signal in active Caspase 3 recombinant protein.
|
|
Cellular localization
|
Cytoplasmic
|
|
Research areas
|
Neuroscience >> Neurology process >> Neurodegenerative disease >> Other
Cell Biology >> Apoptosis >> Intracellular >> Caspases etc >> Caspases
Neuroscience >> Neurology process >> Neurodegenerative disease >> Alzheimer's disease >> Other
|
|
Relevance
|
The 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.
|
|
|
|
|
Database links
|
The links below go to external sites and will open in a new browser window |
|
Raised in
|
Rabbit
|
|
Clonality
|
Polyclonal
|
|
Isotype
|
IgG
|
|
Purity
|
Immunogen affinity purified
|
|
Storage buffer
|
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
|
|
Form
|
Liquid
|
|
Concentration
|
Batch dependent within range: 0.30 - 1.00 mg/ml
|
|
Storage instructions
|
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
|
| |
At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this active Caspase 3 antibody is on this datasheet. But please do contact us if you would like any reassurance! |
 |
|
|
See below for active Caspase 3 antibody images, references, products related to ab13847 and other tools.
|
| |
active Caspase 3 antibody images:
|
 Immunohistochemistry (Frozen sections) - active Caspase 3 antibody (ab13847)
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. Taken from an Abreview submitted by Sophie Pezet
|
| |
Immunocytochemistry/ Immunofluorescence - active Caspase 3 antibody (ab13847)
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. This image is courtesy of an Abreview submitted by Dr Kirk McManus
See Abreview
|
| |
Immunocytochemistry/ Immunofluorescence - active Caspase 3 antibody (ab13847)
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). Roberto Giambruno, Marilena Ciciarello and Patrizia Lavia
|
| |
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - active Caspase 3 antibody (ab13847)
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. This image is courtesy of an Abreview submitted by Dr Michael Levin
See Abreview
|
| |
Western blot - active Caspase 3 antibody (ab13847)
All lanes : 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.
Secondary
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
|
| |
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - active Caspase 3 antibody (ab13847)
ICC/IF image of ab13847 stained MCF7 cells. The cells were 100% methanol fixed (5 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 100% methanol fixed (5 min) Hek293 and HepG2 cells at 5µg/ml.
|
| |
Western blot
All lanes : active Caspase 3 antibody (ab13847) at 1/500 dilution
Lane 1 : Hela whole cell lysate at 20 µg
Lane 2 : A431 whole cell lysate at 20 µg
Lane 3 : Jurkat whole cell lysate at 20 µg
Lane 4 : Hela whole cell lysate at 20 µg with active Caspase 3 peptide (ab13848) at 1 µg/ml
Lane 5 : A431 whole cell lysate at 20 µg with active Caspase 3 peptide (ab13848) at 1 µg/ml
Lane 6 : Jurkat whole cell lysate at 20 µg with active Caspase 3 peptide (ab13848) at 1 µg/ml
Lane 7 : Recombinant Human caspase 3 (active) recombinant protein at 0.1 µg
Lane 8 : Recombinant Human caspase 3 (active) recombinant protein at 0.1 µg with active Caspase 3 peptide (ab13848) at 1 µg/ml
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Predicted band size : 17 kDa
Lanes 1-6: Exposure time 5 min.
Lanes 7-8: Exposure time 30 sec.
ab13847 detects a 17kDa band when tested on recombinant active Caspase 3. This was produced by combining the 17kDa and 12kDa subunits produced separately as recombinant proteins. ab13847 also detects a 17kDa band on lysates from HeLa, A431 and Jurkat cells. These bands can be blocked by the immunising peptide. This strongly suggests that ab13847 is specific for active Caspase 3.
|
| |
References for active Caspase 3 antibody (ab13847)
|
|
This product has been used in: (two most recent references)
Reynolds AD et al. Nitrated {alpha}-synuclein-induced alterations in microglial immunity are regulated by CD4+ T cell subsets. J Immunol 182:4137-49 (2009). ICC/IF, Flow Cyt; Mouse. PubMed: 19299711
Rensen WM et al. RanBP1 downregulation sensitizes cancer cells to taxol in a caspase-3-dependent manner. Oncogene 28:1748-58 (2009). ICC/IF; Human. PubMed: 19270727
See all 12 publication references for this product.
If you publish research using ab13847 please let us know so that we can cite the reference on this datasheet.
|
| |
|
Search PubMed (MEDLINE) for references to active Caspase 3
|
| |
active Caspase 3 antibody - more information
|
Customer reviews (feedback) regarding active Caspase 3 antibody |
Customer FAQs regarding active Caspase 3 antibody |
Protocols for active Caspase 3 antibody |
Price and availability of products related to active Caspase 3 antibody |
| |
active Caspase 3 antibody - related products:
|
|
|
| |
active Caspase 3 antibody - other tools:
|
Contact Abcam with a Technical Enquiry about active Caspase 3 antibody |
| |
All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Pricing information loading
