84-1045-69　Livin (ML-IAP) 100ul　L3180-21

特徴

• Apoptosis or programmed cell death is a fundamental cellular process that is essential for normal tissue development and abnormal growth such as cancer, neurodegeneration, autoimmune diseases, and angiogenesis, etc.
• Apoptosis is driven by specialized proteases known as caspases (Cysteine Aspartase).
• The inhibitor-of-apoptosis protein (IAPs) by virtue of inhibiting caspase activity has widespread anti-apoptotic potential family.
• IAPs are characterized by one or more repeats of a highly conserved ~70 amino acid domain termed the baculoviral IAP repeat (BIR).
• There are at least five human IAP family members, c-IAP1, c-IAP2, XIAP, NAIP, and survivin.
• All of the human IAP family members, with the exception of NAIP, have been shown to interact with specific cysteine proteases, or caspases.
• The caspases are synthesized as inactive zymogen forms which upon apoptotic stimulation are proteolytically processed in a sequential manner into their active heterotetrameric forms.
• The BIR domain is critical motif for anti-apoptotic activity and interaction with caspases.
• Many of the IAPs also contain a RING domain near their COOH-termini that is not required for the anti-apoptotic activity.
• A novel member of the IAPs, termed livin (also known as IAP kidney or KIAP; IAP melanoma or MLIAP), encodes a protein with a single BIR domain and a COOH-terminal RING domain.
• Expression of livin inhibited apoptosis by a number of stimuli.
• Livin can bind to caspases and it could inhibit the proteolytic processing of caspase-9 in vitro.
• Livin gene (chromosome 20q13.3) encodes a 280 amino acid protein.
• It has 25-35% identity with other IAP family members (c-IAP1, c-IAP2, XIAP, NAIP, and surviving).
• At a structural level it was similar to survivin with respect to having just a single BIR domain.
• However, livin did not have a coiled-coil domain like survivin, but rather contained a COOH-terminal RING domain found in c-IAP1, c-IAP2, and XIAP.
• Livin gene has two splicing variants that contain open reading frames of 298 and 280 amino acids and both contained a single copy of baculovirus IAP repeat (BIR) and RING domain.
• Both of the isoforms inhibit TNFa induced apoptosis in Jurkat cells.
• Livin expression is low in adult tissues.
• It is relatively expressed at a higher level in developmental tissues and in many cancer cells.
• Livin was not detectable in most normal adult tissues with the exception of the placenta, but was present in fetal brain and in several cancer cell lines (melanoma-derived cell lines, G361 and SK-Mel29.
• Human Livin (1-280 aa.
• ~32kD) was expressed as his-tag protein in E.
• colli and purified >95%.
• For western blot +ve control, it is supplied in SDS-PAGE sample buffer (reduced).
• Load ~10 ul/lane to visualize with antibodies.
• Applications:Suitable for use in ELISA, Western Blotting.
• Other applications not tested.
• Recommended Dilution:Western Blot: 1:1000-1:5000 for neat serum and 1-10ug/ml for affinity pure antibody using ECL.
• Livin is ~32kD protein (1)
• ELISA: 1:10,000-50,000 for neat serum and 0.5-1ug/ml for affinity pure.
• Control peptide can be used to coat ELISA plates at 1ug/ml.
• Optimal dilutions to be determined by the researcher.
• Storage and Stability:May be stored at 4°C for short-term only.
• For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%).
• Freeze at -20°C.
• Aliquots are stable for at least 12 months at -20°C.
• For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
• Further dilutions can be made in assay buffer.