Matrigel (Matrigel® matrix basement membrane growth factor reduced) was sourced from Corning (New York City, NY, USA). Human recombinant TRAIL was purchased from R&D Systems (Minneapolis, MN, USA). Human recombinant VEGF-A and FGF-2 were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Recombinant human trail
Manufactured by R&D Systems
Sourced in United States, Germany
Recombinant human TRAIL is a cytokine that belongs to the tumor necrosis factor (TNF) superfamily. It is a soluble protein produced in mammalian cells.
Automatically generated - may contain errors
Lab products found in correlation
Cycloheximide (chx), by Merck Group (3 mentions)
Anti actin, by Merck Group (3 mentions)
Z vad fmk, by R&D Systems (3 mentions)
Zvad fmk, by Bachem (3 mentions)
Lcl161, by Novartis (3 mentions)
Mda mb 361, by American Type Culture Collection (2 mentions)
Anti bim, by BD (2 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (2 mentions)
Anti mcl 1, by Santa Cruz Biotechnology (2 mentions)
Anti dr4, by Abcam (2 mentions)
Anti ciap2, by Santa Cruz Biotechnology (2 mentions)
Anti dr5, by Cell Signaling Technology (2 mentions)
Anti bcl xl, by Cell Signaling Technology (2 mentions)
Anti xiap, by BD (2 mentions)
Anti parp, by Cell Signaling Technology (2 mentions)
Z ietd fmk, by Merck Group (2 mentions)
Q vd oph, by Merck Group (2 mentions)
Tg101209, by Selleck Chemicals (2 mentions)
Sd1029, by Merck Group (2 mentions)
Ac lehd cmk, by Merck Group (2 mentions)
25 protocols using recombinant human trail
1
Matrigel and Growth Factor Protocol
2
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)].
3
Sensitizing Cells to TRAIL-Induced Death
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The sensitizing effect was achieved with pre-treatment for 24 h with the bivalent Smac mimetic BV6 (kindly provided by Genentech). Cell death was induced with treatment with human recombinant TRAIL (R&D Systems, Wiesbaden, Germany). Cell death was inhibited with zVAD.fmk (Bachem, Heidelberg, Germany), anti-TRAIL antibody (No. 2E5; Enzo Life Sciences, Lörrach, Germany), zAAD-CMK (Calbiochem Merck, Darmstadt, Germany) and Etanercept (Enbrel, Merck, Darmstadt, Germany).
4
TRAIL-Induced Cell Viability Assay
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The LoVo cells were seeded at a density of 1 × 104 cells/cm2 in 96-well plates and cultured for 24 h. The cells were treated with human recombinant TRAIL (R&D systems), incubated for 24 h, treated with 10 μL of water soluble tetrazolium salt (WST)-1 reagent (Roche, Indianapolis, IN, USA), and incubated for 1 h at standard culture conditions. Then, the absorbance was measured using a microplate reader (Molecular Devices, San Jose CA, USA) at 450 nm.
5
Establishment of TRAIL-Resistant Cell Lines
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In order to secure stable TRAIL-resistant cells, parental LS-LiM6 or DLD1 cells were treated with 100 ng/ml human recombinant TRAIL (R&D Systems, Danvers, MA, #375-TEC) for 24 h. The apoptotic cells were then removed and the surviving cells were propagated in the presence of 10 ng/ml TRAIL for a total of five cycles resulting in stable TRAIL-resistant cell lines LiM6-TR and DLD1-TR. The sensitivity of TRAIL-resistant cell lines to TRAIL was periodically examined by PI (Sigma, St. Louis, MO) staining and flow cytometry, and never exceeded 5% cell death as described before.10 (link)
6
Apoptosis Induction and Inhibition
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For induction of apoptosis the BH3 mimetics A1331852, ABT-199 (Selleck Chemicals, Houston, TX) or S63845 (ApexBio, Houston, TX), and human recombinant TRAIL (R&D Systems, Wiesbaden, Germany) were used. To inhibit cell death zVAD.fmk (Bachem, Heidelberg, Germany) or anti-TRAIL antibody (#2E5, Enzo Life Sciences, Lörrach, Germany) were added to the experiments.
7
miR-26b Modulation of Apoptosis
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miR-26b mimics and negative control oligonucleotide (NC oligo) were purchased from RiboBio Company (China). The sequences of the above RNA oligos were as follows: miR-26b mimics: 5′-UUCAAGUAAUUCAGGAUAGGU-3′; NC oligo: 5′-UGUAAUAAUGGAACUCGGAUU-3′. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), trypan blue, and Annexin V-FITC Apoptosis Detection Kit were obtained from Sigma-Aldrich (USA). Antibodies for rabbit anti-human Mcl-1 and rabbit anti-human β-actin were purchased from Cell Signaling (USA). Human recombinant TRAIL was obtained from R&D Systems (USA).
8
Characterization of Burkitt Lymphoma Cell Lines
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Human BL cell lines RAMOS, DG-75, RAJI, BJAB and DAUDI were obtained from the German Collection of Microorganisms and Cell cultures (DSMZ, Braunschweig, Germany) and BL-2, BL-30, Seraphine, BL-60, BL-70 and Salina cells were kindly provided by T. Oellerich, Department of Medicine II, Hematology/Oncology, University Hospital Frankfurt, Germany. All cell lines were authenticated by STR profiling and continuously monitored for mycoplasma contamination. Cells were cultured in RPMI 1640 (Life Technologies), supplemented with 10% or 20% fetal calf serum (FCS) and 1% penicillin/streptomycin (Invitrogen). The bivalent Smac mimetic BV6 was kindly provided by Genentech. The broad-range caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethylketone (zVAD.fmk) was purchased from Bachem, Necrosulfonamide (NSA) from Toronto Research Chemicals Inc., GSK’872 and Necrostatin-1s (Nec-1s) from Merck and Dabrafenib from Selleck Chemicals. Recombinant human TRAIL was obtained from R&D Systems, human recombinant TNFα from PeproTech and human multimeric FASL from AdipoGen. Doxycycline hydrochloride (DOX) was purchased from Sigma-Aldrich. LCL-161 was purchased from Novartis and AT-406, Birinapant and SGI-110 (Guadecitabine) were obtained from Selleck Chemicals. All other chemicals were purchased from Sigma-Aldrich or Carl Roth unless indicated otherwise.
9
Evaluation of Novel Apoptosis Inducers
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LCL161 was synthesized and provided by Novartis (Basel, Switzerland) under a Material Transfer Agreement (MTA). TG101209 was purchased from Selleckchem (Houston, TX). SD1029 was purchased from EMD Millipore (Billerica, MA). Recombinant human TRAIL and Fas-L were purchased from R&D systems (Minneapolis, MN). Pan-caspase inhibitor Q-VD-OPH, caspase 9 inhibitor Ac-LEHD-CMK and caspase 8 inhibitor Z-IETD-FMK were purchased from EMD Millipore.
10
TRAIL-induced Apoptosis in Kidney Cells
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Mouse kidney cells (TCMK-1) were a gift from Dr. T.J. Lee (Yeungnam University, Korea). Primary culture of human mesangial cells (Cryo NHMC) were purchased from Clonetics (San Diego, CA, USA), and other cell lines are from the ATCC (Manassas, VA, USA). All cells were maintained in Dulbecco’s modified Eagle’s medium, and supplemented with 10% FBS, 5% antibiotic solution, and 100 μg/mL gentamycin. Garcinol and lactacystin was purchased from Enzo Life Sciences (Ann Arbor, MI, USA). Recombinant human TRAIL and z-VAD-fmk were purchased from R&D (Minneapolis, MN, USA). Calbiochem supplied N-acetyl-l -cysteine (NAC), trolox, and MG132 (San Diego, CA, USA). The following antibodies were used: anti-XIAP, -Bcl-2, -Mcl-1, -survivin, -Cbl, and -Itch (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-c-FLIP (ALEXIS Corporation, San Diego, CA, USA); anti-PARP, -DR5, -PSMA5, -PSMD4/S5a, -cleaved caspase-3 (Cell Signaling Technology, Beverly, MA, USA); anti-caspase-3 (Enzo Life Sciences, Ann Arbor, MI, USA); anti-DR4 (Abcam, Cambridge, MA, USA); anti-actin (Sigma-Aldrich, St. Louis, MO, USA). Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Bioneer (Daejeon, Korea) supplied DR5 siRNA and GFP siRNA (control siRNA), respectively. Sigma Chemical Co. supplied other reagents (St. Louis, MO, USA).
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