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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. Mitochondria play a pivotal role in the regulation of programmed cell death or apoptosis. Apoptosis is driven by two classes of specialized proteases known as caspases (Cysteine aspartase). The initiator caspases can be activated by self-cleavage. The effector caspases are then activated in an amplification cascade. The first such factor (Cytochrome-C) to be described binds to a cytoplasmic scaffolding protein called Apaf-1. Binding of the mitochondria factor allows Apaf-1 to form a ternary complex with, and activate, the initiator pro-caspase-9. Active caspase-9 then turns on downstream effector caspases, initiating apoptosis. 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 domains termed the baculoviral IAP repeat (BIR) that is essential for anti-apoptotic activity. 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. C-IAP1, c-IAP2, XIAP, and survivin have been reported to bind to and inhibit the active forms of the terminal caspases-3, -7, and -9 but do not interact with caspases-8, which is the most proximal caspase from the TNF-a/Fas receptor. Another mitochondrial-derived factor, termed Apoptosis inducing factor (AIF), has been identified. AIF is sufficient to induce apoptosis of isolated nuclei. It normally resides in the mitochondrial but translocate to the nucleus. AIF induces mitochondrial to release Cytochrome-c and caspase-9. It has been found in liver and many other tissues.

A new intracellular membrane protein, Aven (from Aventine, a Roman stronghold) has now been shown to bind both Bcl-xl and Apaf-1. Aven is a conserved protein that has broad tissue distribution with prominent expression in heart, skeletal muscle, kidney, liver, pancreas, testis, and several established cell lines (HeLa, IB4, and Raji). Aven interferes with the ability of Apaf-1 to self-associate, and subsequent inhibition of Apaf-1 mediated activation of caspases.

X-chromosome-linked inhibitor of apoptosis protein (XIAP also known as MIHA/IAPA or RIPA-3, MIPA-3 or HLIP) is a potent modulator of apoptosis. XIAP displays 3 tandem BIR domains and C-terminal ring finger motif. A single BIR domain is sufficient for anti-apoptotic activity. XIAP is expressed in most tissues.

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 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).

A small CARD-containing decoy molecule termed ICEBERG regulates activation of caspase-1. This decoy protein binds the corresponding CARD motif of caspase-1, inhibiting and/or displacing the upstream activator RIP2. Structurally, ICEBERG has resemblance to the death-domain-fold superfamily. Iceberg is a 90-aa protein. It is 52% identical to the caspase-1 CARD. It is primarily expressed in the heart and placenta. The related CARD-containing molecules caspase-1 and RIP2 are also expressed in the heart and placenta as well as in numerous other tissues.

Recently, a human gene encoding a structurally and unique IAP designated Survivin has been identified. Survivin contains a single baculovirus IAP repeat and lacks a C-terminal RING finger. It has the property of oncofetal antigens: highly expressed in less-differentiated embryonic cells or rapidly dividing tumor cell but not in fully differentiated adult tissues. Elevated levels of Survivin are found in human fetal lung, liver, heart, kidney, and gastrointestinal tract. In mouse embryonic tissues, Survivin is detected in most tissues. High level of Survivin was found in most common human cancer, including cancers of the lung, colon, pancreas, prostate, and breast. Expression of Survivin also correlated with the presence of both p53 and bcl-2.

Interestingly, Survivin was identified by hybridization screening of human genome libraries with the cDNA of a factor Xa receptor, Effector cell Protease Receptor-1 (EPR-1). Survivin coding strand has significant sequence homology with EPR-1 suggesting a potential for functional interaction between these two proteins. Factor Xa interacts with EPR-1. EPR-1 is expressed in vascular endothelial cells and smooth muscle cells. Survivin and EPR-1 are encoded by structurally and topographically distinct messages from potentially originating from gene cluster at chromosome 17q25. Overexpression of EPR-1 increased apoptosis and inhibited growth of transformed cells.

P53 tumor-suppressor protein functions to inhibit the growth of tumor cells. Several p53-induces genes play a role in the induction of apoptosis. Through global profiling of genes that were expressed soon after p53 expression, a novel gene termed PUMA (p53-upregulated modulator of apoptosis) as a target for p53 activation. PUMA/JFY1 is expressed in most tissues. It is exclusively located in mitochondria and it induces cytochrome-C release. Exogenous expression of PUMA resulted in an extremely rapid and profound apoptosis that occurred much earlier than that resulting from exogenous expression of p53. PUMA is alternatively spliced to produce 2-4 transcripts PUMA alpha (193-aa, ~25 kDa), -beta (131-aa, ~16 kDa), gamma, and delta (101-aa). The gamma and delta forms were not detectable in endogenous cells. PUMA alpha and beta displayed similar apoptosis inducing ability.

Apoptosis occurs not only during programmed cell death, but also during the death process induced by some cytotoxic T cells. A protein ligand, FASL, was identified that triggers cell death by binding to the cell surface receptor variously known as FAS or APT1 family of receptors that includes the 2 tumor necrosis factor (TNF) receptors. The FAS antigen is expressed not only in the cells of the immune system but also in the liver, lung, ovary, and heart, where its function is unclear. FAS ligand/FASL (human 281-aa, ~ 32 kDa, chromosome 1q23) is a type II transmembrane protein that belongs to the tumor necrosis factor family. FASL is expressed in activated splenocytes and thymocytes, consistent with its involvement in T-cell-mediated. FASL is proteolytically cleaved at the cell surface and relased into the extracellualr fluid. FASL is alternative spliced into two forms: Defects in TNFSF6 are a cause of autoimmune lymphoproliferative syndrome (ALPS), also known as Canale-Smith syndrome (CSS), a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly.

    
 

M= Mouse; R=Rat; H=Human; Rb=Rabbit; G=goat; B=Bovine, MO=Monkey; P=pig; CT= near C-terminus; NT=near N-terminus; Internal=Middle of protein. EC=extracellular; CP=cytoplasmic domains *

** Expected antibody crossreactivity information is mostly based upon high (>70%) sequence conservation of antigenic/control peptides in various species. When antibody crossreactivity has actually been experimentally confirmed in various species, it will be mentioned in the appropriate data sheets.

"Neat Antisera or antisera" are the unpurified antiserum and it is suitable for ELISA and Western.
"Affinity pure" IgG may be more suitable for immunohistochemical (IHC) applications and to reduce background in most immunological applications including ELISA and Western.
"Control peptides" can not be used for Western as they are very short peptides. They are intended for ELISA or antibody blocking studies to establish antibody specificity.
Western blot +ve protein controls, where available, are semi-pure, pure or recombinant proteins that are formulated in SDS-PAGE sample buffer. They are recommended to be used for Western (load 10 ul/lane) for visulaization with antibodies.

 

List of Publications using ADI's Antibodies of various apoptosis related products

AIF
Dirks AJ 2003 AGING AND LIFELONG CALORIE RESTRICTION RESULT IN ADAPTATIONS OF SKELETAL MUSCLE APOPTOSIS REPRESSOR, APOPTOSIS-INDUCING FACTOR, X-LINKED INHIBITOR OF APOPTOSIS, CASPASE-3, AND CASPASE-12 Free Radical Biology and Medicine WB rat.

Livin
Tu SP 2003 Suppression of Survivin Expression Inhibits in Vivo Tumorigenicity and Angiogenesis in Gastric Cancer Cancer Res., Nov 2003; 63: 7724 - 7732. WB IHC gastric epithelial cancer cells

Livin
ka H 2003 Temporal and Spatial Patterns of Expression of Inhibitors of Apoptosis in Human Placentas Am. J. Pathol., Aug 2003; 163: 413 - 422 Wb IHC human placenta, -ve results for WB, nuclear staining

Parkin
Hase A et al 2002 Characterization of parkin in bovine peripheral nerve Brain Res. 930, 143-149 WB, IHC, also ab from chemicon' human brain,

Survivin
Sasaki S et al 2002 Expression of survivin, an inhibitor of apoptosis protein, in tumors of the nervous system Acta Neuropathologica IHC,, human brain formalin-fixed, paraffin embedded

Survivin
Tu SP 2003 Suppression of Survivin Expression Inhibits in Vivo Tumorigenicity and Angiogenesis in Gastric Cancer Cancer Res., Nov 2003; 63: 7724 - 7732. WB IHC gastric epithelial cancer cells

Survivin
Nakagawa Y 2003 Differential expression of survivin in bone marrow cells from patients with acute lymphocytic leukemia and chronic lymphocytic leukemia Leukemia Research, In Press IHC bone marrow tumor

Survivin
Mohapatra S 2003 Roscovitine Inhibits STAT5 Activity and Induces Apoptosis in the Human Leukemia Virus Type 1-Transformed Cell Line MT-2 Cancer Res., Dec 2003; 63: 8523 - 8530 WB iap1/2 from transduction

Survivin
Tran, Jennifer 1999 Marked induction of the IAP family anti-apoptotic proteins survivin and XIAP and VEGF in vascular endothelial cells BBRC 264, 781-788
Survivin
Okada E et al 2001 Survivin expression in tumore cell nuclei is a function of favorable prognosis in gastric cancer patients Cancer Lett. 163, 109-116 WB, IHC, IF formalin-fixed, paraffin embedded

Survivin
Chantalat L 2000 Crystal Structure of Human Survivin Reveals a Bow Tie&hibar;Shaped Dimer with Two Unusual -Helical Extensions Molecular Cell, 6, 183-189 WB, IF

Survivin
Kanwar, Jagat R. 2001 Effects of Survivin Antagonists on Growth of Established Tumors and B7-1 Immunogene Therapy J Natl Cancer Inst 93: 1541-1552 WB, IHC, flow mouse tumor

Survivin
Guo F et al 2002 Ectopic overexpression of second mitochondria-derived activator of caspases (Smac/DIABLO) or cotreatment with N-terminus of Smac/DIABLO peptide potentiates epothilone B derivative-(BMS 247550) and Apo-2L/TRAIL-induced apoptosis Blood 99, 3419-3429 WB IHC MAb survivin Jurkat T-cell leukemia and SKW6.4 B lymphoblast cells

Survivin
Krieg A 2002 Expression of different survivin variants in gastric carcinomas: first clues to a role of survivin-2B in tumour progression British Journal of Cancer 86, 737 - 743 WB gastric carcinoma/variants

Survivin
Ikeguchi M 2002 Expression of Survivin Messenger RNA Correlates With Poor Prognosis in Patients With Hepatocellular Carcinoma Diagnostic Molecular Pathology 2002;11:33-40

Survivin
nakanishi K 2002 survival in patients with transitional cell carcinoma of the upper urinary tract Virchows Archiv 2002 in press IHC paraffin sections/autoclave/urinary bladder tumors

Survivin
Ikeguchi M 2002 Inducible Nitric Oxide Synthase and Survivin Messenger RNA Expression in Hepatocellular Carcinoma Clin Cancer Res. 8, 3131-3136 WB, human tumors/aslo I-NOS from chemic/ADI

Survivin
Wittmann S 2003 Flavopiridol Down-Regulates Antiapoptotic Proteins and Sensitizes Human Breast Cancer Cells to Epothilone B-induced Apoptosis Cancer Res., Jan 2003; 63: 93 - 99 WB, MB-468 and SKBR-3

Survivin
Yamamoto T 2002 Downregulation of survivin expression by induction of the effector cell protease receptor-1 reduces tumor growth potential and results in an increased sensitivity to anticancer agents in human colon cancer European Journal of Cancer, 38, 2316-2324 WB, IHC pf/paraffin HT29 human colon adenocarcinoma cells

Survivin
Griffin D 2003 Molecular determinants of epothilone B derivative (BMS 247550) and Apo-2L/TRAIL-induced apoptosis of human ovarian cancer cells Gynecologic Oncology, Volume 89, Issue 1, April 2003, Pages 37-47 WB human epithelial ovarian tumor cell line 2008

Survivin
Shigeno M 2003 Interferon- sensitizes human hepatoma cells to TRAIL-induced apoptosis through DR5 upregulation and NF-B inactivation Oncogene 22, 1653 - 1662 WB Human HepG2, Hep3B and PLC/PRF/5 cell lines

Survivin
nakanishi K 2002 Expression of survivin does not predict survival in patients with transitional cell carcinoma of the upper urinary tract Virchows Archiv 2002 in press IHC paraffin sections/autoclave/urinary bladder tumors

Survivin
Nimmanapalli R 2003 Regulation of 17-AAG-induced apoptosis: role of Bcl-2, Bcl-xL, and Bax downstream of 17-AAG-mediated downregulation of Akt, Raf-1 and Src kinases Blood, Mar 2003; 102: 269 - 275 WB Moab

Survivin
Ikeguchi M 2003 survivin messenger RNA expression is a good prognostic biomarker for oesophageal carcinoma British Journal of Cancer 87, 883 - 887 IHC human tumors/

Survivin
Shankar, Sai Latha 2001 Survivin inhibition induces human neural tumor cell death through caspase-independent and -dependent pathways Journal of Neurochemistry Volume 79, Issue 2, Page 426 WB, msn cells/surv from R&D,
Survivin
Hiroshima K 2002 Proliferative Activity and Apoptosis in Thymic Epithelial Neoplasms Mod. Pathol., Dec 2002; 15: 1326 - 1332

 

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