Western blot analysis of Caspase 1 was performed by loading 35 µg of THP-1 lysate in reducing sample buffer onto a 4–15% Precast Protein Gels. Proteins were transferred to PVDF and then blocked in blocking buffer (TBST+5% non-fat milk) for one hour at room temperature. Caspase 1 was detected at approximately 45 kDa using a caspase 1 polyclonal antibody (Product # PA5-17570) at a dilution of 1:1000 overnight at 4°C on a rocking platform, followed by a Rabbit IgG Horseradish Peroxidase-conjugated Antibody at a dilution of 1:1,000 for one hour. Chemiluminescent detection was performed using SuperSignal™ West Pico Chemiluminescent Substrate (Product # 34080).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues within the p20 subunit of human caspase-1|
|Purification||Antigen affinity chromatography|
|Storage buffer||0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/ml BSA, 50% glycerol|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50|
|Western Blot (WB)||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.
PA5-17570 detects Caspase 1 in WB application with human samples.
It is not recommended to aliquot this antibody.
This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce 2 subunits, large and small, that dimerize to form the active enzyme. This gene was identified by its ability to proteolytically cleave and activate the inactive precursor of interleukin-1, a cytokine involved in the processes such as inflammation, septic shock, and wound healing. This gene has been shown to induce cell apoptosis and may function in various developmental stages. Studies of a similar gene in mouse suggest a role in the pathogenesis of Huntington disease. Alternative splicing of this gene results in five transcript variants encoding distinct isoforms.
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