Product nameAnti-Caspase-3 antibody [E87] (Biotin)
See all Caspase-3 primary antibodies
DescriptionRabbit monoclonal [E87] to Caspase-3 (Biotin)
Tested applicationsSuitable for: WB, IHC-Pmore details
Species reactivityReacts with: Human, Recombinant fragment
Predicted to work with: Rat
Synthetic peptide within Human Caspase-3 aa 50-150. The exact sequence is proprietary.
Database link: P42574
- WB: Active Caspase-3 recombinant protein. IHC-P: Human normal tonsil (FFPE sections)
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab195905 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000. Detects a band of approximately 15 kDa (predicted molecular weight: 31 kDa).|
|IHC-P||1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionInvolved 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.
Tissue specificityHighly 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 similaritiesBelongs to the peptidase C14A family.
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.
- Information by UniProt
- A830040C14Rik antibody
- Apopain antibody
- CASP 3 antibody
IHC image of Caspase-3 staining in a section of formalin-fixed paraffin-embedded Human normal tonsil*, performed on a Leica BondTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab195905, 1/50 dilution, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Anti-Caspase-3 antibody [E87] (Biotin) (ab195905) at 1/10000 dilution +
Recombinant human Cleaved Caspase-3 protein (Active) (ab52101) at 0.1 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab195905 overnight at 4°C. Antibody binding to the target was detected using a HRP conjugated streptavidin amplification step, and visualised using ECL development solution ab133406.
ab195905 has not yet been referenced specifically in any publications.