Immunohistochemistry analysis of Caspase-3 showing staining in the cytoplasm of paraffin-embedded human lung adenocarcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Caspase-3 monoclonal antibody (437800) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Horse, Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant protein derived from the N-terminus of human Caspase 3 protein|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
* 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.
Caspase 3 is ubiquitously expressed and like other caspases is synthesized as an inactive (32kDa) proenzyme. Upon activation, Caspase 3 is cleaved at Asp28-Ser29 and Asp175-Ser176 thereby generating two subunits of 17kDa and 12kDa, respectively. Recent studies have implicated that Caspase 3 is associated with the induction of apoptosis. Activation of Caspase 3 occurs in response to variety of apoptotic inducers including Fas mediated apoptosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.