FAIM-L overexpression, XIAP down-regulation, and their cross-regulation were tested by immunocytochemistry in infected hippocampal primary cultures. Immunocytochemistry was performed using anti-XIAP (1:500; BD Biosciences), anti-FAIM-L (1:500; in house), and rabbit anti-GFP (1:500; Invitrogen), followed by Alexa 568 and Alexa 488-coupled antibodies (1:500; Invitrogen). In DRG neurons, XIAP, FAIM-L, pan-ERK, NF66 and caspase-3 levels were assessed by Western blot using anti-XIAP (BD Biosciences; 1:20,000), anti-FAIM-L (in house; 1:2,000), anti-pan-ERK (BD Biosciences; 1:20,000), anti-NF66 (Covance; 1:10,000), anti-caspase-3 (Cell Signaling; 1:1,000) and Histone H3 (Thermo Scientific; 1:40,000) antibodies.
Anti xiap
Manufactured by BD
Sourced in United States
Anti-XIAP is a laboratory reagent that binds to and inhibits the activity of the XIAP (X-linked Inhibitor of Apoptosis Protein) protein. XIAP plays a role in regulating cell apoptosis, or programmed cell death. Anti-XIAP can be used in research applications to study the mechanisms of cell death and apoptosis.
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Lab products found in correlation
Anti mcl 1, by Santa Cruz Biotechnology (7 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (7 mentions)
Anti ciap2, by Santa Cruz Biotechnology (7 mentions)
Anti parp, by Cell Signaling Technology (7 mentions)
Anti bim, by BD (6 mentions)
Anti dr5, by Cell Signaling Technology (5 mentions)
Anti bcl xl, by Cell Signaling Technology (5 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (5 mentions)
Anti c flip, by Enzo Life Sciences (5 mentions)
Anti caspase 3, by Cell Signaling Technology (4 mentions)
Anti actin, by Merck Group (4 mentions)
Anti dr4, by Abcam (4 mentions)
Anti caspase 8, by Cell Signaling Technology (3 mentions)
Anti bax, by BD (3 mentions)
Z vad fmk, by R&D Systems (3 mentions)
Anti survivin, by R&D Systems (3 mentions)
Cycloheximide (chx), by Merck Group (3 mentions)
Lactacystin, by Enzo Life Sciences (3 mentions)
Anti pro caspase 3, by Enzo Life Sciences (3 mentions)
Bca kit, by Thermo Fisher Scientific (3 mentions)
17 protocols using anti xiap
1
FAIM-L and XIAP Cross-Regulation in Neurons
2
Antibody and Chemical Sources for Cell Signaling
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The antibodies and chemicals were obtained from the following resources; anti-PARP (#556362), anti-XIAP (#610716), anti-FADD (#610399), anti-RIP1 (#610459), anti-p53 (#554147) and anti-p21 (#556430) antibodies (BD Biosciences, San Diego, CA, USA); anti-caspase-3 (#9662), anti-caspase-8 (#9746), anti-caspase-9 (#9508) and anti-Bid (#2002) antibodies (Cell signalling Technology, Beverly, MA, USA); anti-caspase-10 (M059-3) antibody (MBL, WOBURN, MA, USA); anti-Bcl-XS/L (sc-271121), anti-survivin (sc-17779), anti-TRAF2 (sc-876), anti-GFP (sc-9996) and anti-HA (sc-805) antibodies (Santa Cruz, CA, USA); anti-c-FLIP (ALX-804-961) antibody (Enzo Life Sciences, Farmingdale, NY, USA); anti-cIAP1/2 (#07-759) antibody (Upstate Biotech, Waltham, MA, USA); anti-DR5 (#ab181846) antibody (Abcam, Cambridge, UK); anti-DR4 (NB100-56528) antibody (Novus, Centennial, CO, USA); anti-TurboGFP (PA5-22688) antibody (Thermo scientific, Waltham, Massachusetts, USA); anti-actin (A2066), anti-flag (F3165, 1:2,000 dilution) antibodies (Sigma-Aldrich, St. Louis, MO, USA). the pan caspase inhibitor Z-VAD-FMK, MG-132, TPCA-1 (Calbiochem, San Diego, CA, USA); recombinant TNF (R & D Systems, Minneapolis, MN, USA); recombinant human TRAIL/Apo2 ligand (Peprotech, Rocky Hill, NJ, USA).
3
Antibodies and Reagents for Cell Signaling
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Anti-LEFTY, anti-Smad2, and anti-phospho-Smad2 at serine 255 (pSmad2) antibodies were purchased from Abcam (Cambridge, MA, USA). Anti- XIAP, anti-bax, anti-β-catenin, anti-HNF-1β, and anti-p27Kip1 antibodies were bought from BD Biosciences (San Jose, CA, USA). Anti-p21waf1, anti-cyclin D1, anti-p53, anti-bcl2, and anti-Ki-67 antibodies were purchased from Dako (Copenhagen, Denmark). Anti-cyclin A, and anti-cleaved caspase 3 antibodies were from Novocastra (Newcastle, UK), and Cell Signaling Technology (Danvers, MA, USA), respectively. CDDP and the anti-β-actin antibody were purchased from Sigma-Aldrich Chemicals (St. Louis, MO, USA). Recombinant transforming growth factor (TGF)-β1 was purchased from R&D Systems (Minneapolis, MN, USA).
4
Antibodies for Western Blot Analysis
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Western blot analysis was performed as described previously [17] (link) using the following antibodies: anti-cIAP1 (R&D Systems, Inc., Wiesbaden-Nordenstadt, Germany), anti-cIAP2 (Epitomics, Burlingam, CA), anti-XIAP from BD Biosciences, anti–phospho-p65 from Cell Signaling (Beverly, MA), anti-phospho-IκBα and anti-IκBα (Cell Signaling), anti-β-actin (Sigma), anti-p65, and anti-p52 from Santa Cruz Biotechnology (Santa Cruz, CA). Donkey anti-mouse IgG, donkey anti-rabbit IgG, or donkey anti-goat IgG labeled with IRDye infrared dyes was used for fluorescence detection at 680 or 800 nm (LI-COR Biotechnology, Bad Homburg, Germany).
5
Antibody-Based Protein Detection Protocol
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Antibodies against β-Actin, Bcl2, p-ERK, ERK, LC3, p62, XIAP, ATG5 and PARP were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-XIAP and anti-pP27 were purchased from BD Biosciences (San Jose, CA, USA). Anti-Caspase-3 (Active) antibody was purchased from Millipore (Temecula, CA, USA). An anti-ARHI murine monoclonal antibody (ID8) was generated in our laboratory. Doxycycline hyclate (DOX), chloroquine diphosphate salt (CQ), cis-diammine-platinum (II) dichloride (Cisplatin), Z-VAD, Nec-1 and N-acetyl-L -cysteine were purchased from Sigma-Aldrich (St. Louis, MO, USA).
6
Investigating Cancer Cell Signaling Pathways
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Human renal carcinoma (Caki-1 and A498), human lung cancer (A549), and human breast cancer (MDA-MB361) were procured from American Type Culture Collection (Manassas, VA, USA). Human recombinant TRAIL, zVAD-fmk, and anti-survivin were provided by the R&D system (Minneapolis, MN, USA). MG132, PD98059, AG-490, compound C, and NVP-BEZ235 were supplied from Calbiochem (San Diego, CA, USA). Dexamethasone, cycloheximide, AR-A014418, PP242, BAY11-7082, rapamycin, and anti-actin were provided from Sigma Chemical Co. (St. Louis, MO, USA). Anti-PARP, anti-Bcl-xL, anti-DR5, anti-cIAP1, anti-caspase-8, anti-phospho-GSK3β, and anti-GSK3β were provided by Cell Signaling Technology (Beverly, MA, USA). Anti-Bim, anti-Bax, and anti-XIAP were obtained from BD Biosciences (San Jose, CA, USA). Anti-Mcl-1, anti-Bcl-2, anti-cIAP2, and anti-Cbl were purchased from Santa Cruz Biotechnology (St. Louis, MO, USA). SB203580, SP600125, and anti-c-FLIP(L) were obtained from Enzo Life Sciences (San Diego, CA, USA). Anti-DR4 were obtained from Abcam (Cambridge, MA, USA). pCMV-Myc-Cbl plasmid was a gift from Dr. S. J. Kim (CHA University, Korea). GSK3betaS9A (1016) was a gift from Scott Friedman (Addgene plasmid # 49492; http://n2t.net/addgene:49492 ; RRID: Addgene_49492) [36 (link)].
7
Molecular Mechanisms of TRAIL-Induced Apoptosis in Cancer Cells
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Selleckchem supplied R428 and cisplatin (Houston, TX, USA), and R&D system supplied recombinant human recombinant TRAIL and z-VAD-fmk (Minneapolis, MN, USA). Sigma Chemical Co. provided MG132, cycloheximide, carboplatin, oxaliplatin, doxorubicin, and 5-FU (St. Louis, MO, USA). The primary antibodies were as follows: Anti-phospho-Axl (Y702) (Cell Signaling Technology, Beverly, MA, USA), anti-pro-caspase-3 (Cell Signaling Technology), anti-cleaved caspase-3 (Cell Signaling Technology), anti-PARP (Cell Signaling Technology), anti-Bcl-xL (Cell Signaling Technology), anti-DR5 (Cell Signaling Technology), anti-actin (Sigma Chemical Co.), anti-Axl (Santa Cruz Biotechnology, St. Louis, MO, USA), anti-Mcl-1 (Santa Cruz Biotechnology), anti-Bcl-2 (Santa Cruz Biotechnology), anti-cIAP2 (Santa Cruz Biotechnology), anti-Bim (BD Biosciences, San Jose, CA, USA), anti-XIAP (BD Biosciences), anti-survivin (R&D system, Minneapolis, MN, USA), anti-DR4 (Abcam, Cambridge, MA, USA), and anti-c-FLIP (Enzo Life Sciences, San Diego, CA, USA). The siRNAs were as follows: GFP (control) siRNA (Bioneer, Daejeon, Korea), Axl siRNA (Santa Cruz Biotechnology), and DR5 siRNA (Invitrogen).
8
Protein Expression Analysis Protocol
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The total cell lysates were prepared by resuspending 0.45×106 cells in 20–50 µl of RIPA lysis buffer (50 mM Tris buffer, 20 mM HEPES, 100 mM NaF; 120 mM NaCl, 0.5% Triton X-100, 100 µM Na3VO4, pH 7.6) Total lysates was quantified using a BCA kit (#23225; Thermo Fisher Scientific) according to the manufacturer's protocols. The proteins (30–70 µg) were isolated using 10 or 12% SDS-PAGE gels and electrotransferred onto NC membranes (GE Healthcare). Target proteins were identified using the respective antibodies and Immobilon Western Chemiluminescent HRP Substrate Solution (WBKLS0100; Millipore) and visualized by Davinch-Chemi (CAS-400SM; Davinch-K). Anti-PARP antibody (1:1,000; #9542) and anti-Bax (1:1,000; #2772) were obtain from Cell Signaling Technology. Anti-caspase-3 (1:3,000; ADI-AAP-113) and anti-FLIP (1:700; ALX-804-961-0100) were purchased from Enzo Life Sciences. Anti-Bcl-2 (1:700; sc-7832), anti-Bcl-xL (1:1,000, sc-634), anti-Mcl-1 (1:1,000; sc-12756), anti-cIAP1 (1:1,000; sc-7943), anti-cIAP2 (1:1,000; sc-517317) and anti-β-actin antibody (1:5,000; sc-47778) were supplied by Santa Cruz Biotechnology, and anti-XIAP (1:10,000; 610717) antibody was obtained from BD Biosciences.
9
Comprehensive Western Blot Analysis of Cell Death Signaling
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Western blot was performed as described [21 (link)]. Primary antibodies used were anti-actin (1:2000 MP Biomedicals C4), anti-caspase-8 (1:300 Cell Signaling #9746 and 1:300, Abcam ab32125), anti-caspase-3 (1:1000 Cell Signaling #9662), anti-caspase-7 (1:400 Cell Signaling #12827), anti-RIP1 (1:500 BD Biosciences 610458), anti-XIAP (1:400 BD Biosciences 610762), anti-cIAP1 (1:200 R&D Systems AF8181), anti-p100/p52 (1:500 Cell Signaling #4882) and anti-c-FLIP (1:400 Cell Signaling #5634). Secondary horseradish peroxidase-labeled antibodies were from GE Healthcare and Dako and were used at 1:5000 dilutions. For the chemiluminescent reaction, Supersignal Substrate (Thermo Scientific) was used. Chemiluminescence was detected with a LAS-1000 CCD camera and Image Reader LAS-1000 Pro v2.6 software (Fujifilm, Tokyo, Japan) or an Amersham Imager 600 (GE Healthcare, Chicago, IL, USA).
10
Hispolon sensitizes TRAIL-induced apoptosis
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American Type Culture Collection (Manassas, VA, USA) supplied the all cells and the transformed mouse kidney cells (TCMK-1) was a gift from Dr. TJ Lee (Yeungnam University, Korea). The cells were cultured using Dulbecco’s modified Eagle’s medium containing 10% FBS, 20 mM HEPES buffer and 100 μg/mL gentamycin. Hispolon was purchased from ENZO life Science (Farmingdale, NY, USA). The recombinant human TRAIL was purchased from KOMA Biotech (Seoul, Korea). Anti-DR5 (1:700, #8074), Bcl-xL (1:700, #2764), and anti-PARP (1:700, #9542) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-DR4 (1:1,000, ab8414), and anti-cIAP1 (1:700, ab154525), antibodies were purchased from Abcam (Cambridge, UK). Anti-c-FLIP (1:700, ALX-804-961-0100) antibody was obtained from Enzo Life Sciences. Anti-Mcl-1 (1:700, sc-819) and anti-cIAP2 (1:700, sc7944) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-XIAP (1:1,000, 610762) antibody and anti-Bim (1:700, AB17003) antibodies were purchased from BD Biosciences (San Jose, CA, USA). Cyclohexamide and other reagents were purchased from Sigma Chemical Co. (St. Louis, MO, USA).
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