Preparation And Storage
Recommended Assay Procedures
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Companion Products
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Apoptosis, a selective process of genetically programmed cell death, occurs during normal cellular differentiation and development of multicellular organisms. Apoptotic cells are characterized by loss of cell volume, plasma membrane blebbing, nuclear condensation, chromatin aggregation, and endonucleocytic degradation of DNA into nucleosomal fragments. Caspase-3 (CPP32, Yama, apopain) is the most extensively studied apoptotic protein. An apoptotic signal induces the intracellular cleavage of caspase-3 from an inactive proform to an active fragment. The active form of caspase-3 cleaves several other apoptotic proteins including DNA Fragmentation Factor (DFF). DFF is thought to be either directly or indirectly responsible for the DNA laddering that is a hallmark of apoptosis. It is a heterodimer of 40 kDa (DFF40) and 45 kDa (DFF45) subunits. Cleavage of DFF45 into three fragments is a prerequisite for DFF-mediated DNA fragmentation. Identity of the DFF fragment that mediates DNA laddering remains unclear. However, it is clear that DFF functions downstream of caspase-3 and is essential for the characteristic fragmentation of DNA during apoptosis.
Development References (5)
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Fulda S, Meyer E, Debatin KM. Inhibition of TRAIL-induced apoptosis by Bcl-2 overexpression. Oncogene. 2002; 21(15):2283-2294. (Biology: Western blot). View Reference
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Inohara N, Koseki T, Chen S, Wu X, Nunez G. CIDE, a novel family of cell death activators with homology to the 45 kDa subunit of the DNA fragmentation factor. EMBO J. 1998; 95(9):2526-2533. (Biology). View Reference
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Li J, Chen P, Sinogeeva N, et al. Arsenic trioxide promotes histone H3 phosphoacetylation at the chromatin of CASPASE-10 in acute promyelocytic leukemia cells. J Biol Chem. 2002; 277(51):49504-49510. (Biology: Western blot). View Reference
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Liu X, Li P, Widlak P. The 40-kDa subunit of DNA fragmentation factor induces DNA fragmentation and chromatin condensation during apoptosis. Proc Natl Acad Sci U S A. 1998; 95(15):8461-8466. (Biology). View Reference
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Liu X, Zou H, Slaughter C, Wang X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell. 1997; 89(2):175-184. (Biology). View Reference
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