Livin Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Livin (BIRC7/KIAP) is a member of the family of inhibitor of apoptosis proteins (IAP) and contains a single copy of a baculovirus IAP repeat (BIR) as well as a RING-type zinc finger domain (reviewed in Ma et al 2006 and Crnkovic-Mertens et al 2006). Resistance towards apoptosis is a hallmark of cancer cells, and overexpression of IAPs can contribute to the development of cancer though inhibiting apoptosis (reviewed in Wright et al. 2005). IAP proteins inhibit apoptosis by binding to and inhibiting caspases through their BIR domain(s). Some IAP members, like Liven, also have a RING-type finger motif at their carboxyl-terminal which may enhance anti-apoptotic activity. Many RING finger-containing IAPs possess E3 ubiquitin ligase activity, and it has been shown that Liven acts an E3 ubiquitin ligase for targeting the degradation of Smac/DIABLO. Smac/DIABLO functions by inhibiting IAP-caspase interactions, thereby promoting apoptosis. Thus degradation of Smac/DIABLO is thought to be a mechanism to enhance Livin anti-apoptotic activity, thereby promoting cell survival. Two splicing variants of Livin, alpha and beta, have been identified. The two isoforms are thought to have different anti-apoptotic properties; however, there are conflicting reports are to whether they actually differ in their biological activities. There is accumulating evidence that Liven plays a significant role in a spectrum of tumor types and it is thought that Liven may have potential as a diagnostic and prognostic tumor marker. Liven is expressed at low levels in adult tissues, and relatively higher levels in developmental tissues and in many cancer cells. Certain cancer patients develop autoantibodies against liven and in a number of cancers, Liven is detected only in the tumors but not, or to substantially lower levels, in the corresponding normal adjacent tissue. This antibody recognizes Livin, both isoform alpha and isoform beta. Human Livin isoform alpha is a 298 amino acid (aa) protein, GenBank no. NP_647478. Human Livin isoform beta is a 280 aa protien, GenBank no. NP_071444.1. The antibody will also recognize other isoforms of Livin that contain the peptide immunogen sequence.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-06455

• Livin Antibodies
• Livin ELISA Kit
• Livin Lysates
• Livin Proteins
• Livin Proteins and Enzyme