Description
The caspase family of cysteine proteases plays a key role in apoptosis and inflammation. ICE (interleukin-1ß converting enzyme) was the first member of this family to be discovered following a search for human proteins with homology to ced-3, a cell death gene identified in C. elegans. "Caspase" has been adopted as a root name for all family members based on the properties of these enzymes. The "c" reflects a cysteine protease mechanism and "aspase" refers to their ability to cleave after aspartic acid residues. Caspases are synthesized as inactive proenzymes that are processed in cells undergoing apoptosis by self-proteolysis and/or cleavage by another protease. The processed forms consist of large (17-22 kDa) and small (10-12 kDa) subunits which associate to form an active enzyme (34 kDa). Caspase-6 exists as two isoforms, designated Mch2α (caspase-6) and Mch2ß, a proteolytically inactive isoform which lacks half of the large subunit. Caspase-6 is most closely related to caspase-3, and can be proteolytically cleaved/activated by caspase-3 into large (p18) and small (p11) subunits. Similarly, caspase-3 has been shown to be activated by caspase-6, suggesting that these proteases may generate an amplification cycle during apoptosis. Other proteolytical targets are shown below in the Western blot analysis of caspase-6 (lane 1): Lamin A and a cytosolic nuclease which translocates to the Daudi B cell lysate after probing with anti-nucleus to promote DNA fragmentation. Caspase-6 has also been shown to cleave Fak (focal adhesion kinase), a non-receptor protein tyrosine kinase that transduces cell signals. Clone B93-4 identifies full length caspase-6 at survival and proliferation signals from contact sites between the cell surface and extracellular matrix. In this instance, caspase-6 cleavage inhibits the anti-apoptotic function of Fak. Therefore, the protease activity of caspase-6 plays both activating and inhibitory roles in apoptotic pathways. Caspase-6 is observed at ~34 kDa in SDS-PAGE. The antibody detects both full length caspase-6 (34 kDa), as well as the p11 subunit of the active enzyme. A synthetic peptide sequence corresponding to amino acids 271-285 (KKQVPCFASMLTKK) of human caspase-6 was used as immunogen.
Format
- FormatPurified
Suggested Companion Products
HRP Goat Anti-Mouse Ig RUO 1mLCat No: 554002
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Preparation and Storage
Store undiluted at 4°C.The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Applications include western blot analysis (2 µg/ml). Recombinant human caspase-6 or 293 human embryonic kidney cells (ATCC CRL-1573) are suggested as positive controls for this application.
