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Immunofluorescent analysis of Caspase-3 was performed on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0. 25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ Caspase-3 recombinant rabbit oligoclonal antibody (Product # 710431) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381) Panels d is a merged image showing cytoplasmic localization and panel e a is no primary antibody control. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 30–43 of human Casp3|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:100-1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies - an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Caspases are a family of cysteine proteases that centrally controls apoptotic machinery. Caspases can be grouped according to their substrate specificities that are largely determined by the amino acids preceding the cleavage site. One group of caspases that include -6, -8, is specific for the substrate V/LEXD. This substrate is a site similar to those found in caspase proenzymes. This group of caspases may function as initiators of a proteolytic cascade by activating pro-caspases to amplify a death signal. A second group of caspases is specific for the substrate DEXD that is related to sites found on target proteins cleaved during apoptosis.
Caspase-3 is a member of the interleukin-1 beta converting enzyme, Caspase-8, nuclear lamins and others. The overexpression of Caspase-3 can result in apoptosis. Likewise, the inhibition of Caspase-3 or other caspases can prevent cells from entering the apoptotic pathway. Recent evidence has revealed a link between plasma caspase-3 and atherosclerosis and it role in activation of apoptosis in breast cancer mediated by siRNA-mediated Apollon silencing. This antibody is specific for the cleaved (active) form of caspase-3.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
apopain; apoptosis-related cysteine protease; CASP-3; caspase 3; caspase 3, apoptosis-related cysteine peptidase; caspase 3, apoptosis-related cysteine protease; caspase-3; CPP-32; cysteine protease CPP32; PARP cleavage protease; procaspase3; protein Yama; SREBP cleavage activity 1; Yama
CASP3; CPP32; CPP32B; SCA-1