The human gastric cancer cell line MGC-803 was purchased from American Type Culture Collection (Manassas, VA, USA); the RPMI-1640 medium, fetal bovine serum (FBS) and polyvinylidene fluoride (PVDF) membrane were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany); the primary antibodies [B cell lymphoma (Bcl)-2, Bcl-2 Associated X, Apoptosis Regulator (Bax), tumor protein P73 (TP73) and GAPDH] and the secondary antibody were purchased from Cell Signaling Technology Inc., (Danvers, MA, USA); the MTT assay kit was purchased from Eli Lilly and Company (Indianopolis, IN, USA); the miR647 mimic/inhibitor were purchased from Sigma-Aldrich; Merck KGaA, and the cell transfection kit were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
B cell lymphoma 2
Manufactured by Cell Signaling Technology
Sourced in United States
Bcl-2 is a protein that plays a key role in regulating apoptosis, or programmed cell death. It is a member of the Bcl-2 family of proteins and functions as an anti-apoptotic factor, helping to prevent cell death.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Cell Signaling Technology (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Mgc 803, by American Type Culture Collection (1 mentions)
Phospho p53, by Cell Signaling Technology (1 mentions)
Antibodies against cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Phospho bcl 2, by Cell Signaling Technology (1 mentions)
Pr 619, by MedChemExpress (1 mentions)
Caspase 4, by Cell Signaling Technology (1 mentions)
Jnk sc 571, by Santa Cruz Biotechnology (1 mentions)
C myc, by Cell Signaling Technology (1 mentions)
Cleaved caspase 8, by Cell Signaling Technology (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Rhodamine 123, by Merck Group (1 mentions)
Anti nitrotyrosine nt, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Phosphorylated ampkα, by Cell Signaling Technology (1 mentions)
Fluo 4 acetoxymethyl ester fluo 4, by Thermo Fisher Scientific (1 mentions)
Phosphorylated acetyl coa carboxylase, by Cell Signaling Technology (1 mentions)
Anti 4 hydroxynonenal 4 hne, by Merck Group (1 mentions)
4 protocols using b cell lymphoma 2
1
Investigating miR-647 in Gastric Cancer
2
Delineating Apoptotic Signaling Pathways
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PR-619 was obtained from MedChemExpress (Junction, NJ, USA) and cisplatin from Merck Millipore (Billerica, MA, USA). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA) or Merck Millipore. For Western blot analysis, antibodies against cleaved caspase-3 (#9661), cleaved caspase-8 (#9496), B cell lymphoma (Bcl)-2 (#15071), phospho-Bcl-2 (#2824), phospho-p53 (#9284), caspase-4 (#4450), phospho-Janus kinase (JNK) (#9255), and c-Myc (#18683) were procured from Cell Signaling Technology (Danvers, MA, USA). The antibodies against β-actin (#109) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, #100118) were supplied by GeneTex (Irvine, CA, USA); those against α-tubulin (sc-5286), P21 (sc-6264), P27 (sc-1641), and JNK (sc-571) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). For IHC, the USP14 antibody (#MA5-32821) was purchased from Invitrogen, the USP21 antibody (#17856-1-AP) was purchased from Proteintech (Chicago, IL, USA), and the c-Myc (#ab32072) antibody was purchased from Abcam (Cambridge, UK).
3
Compound C Inhibits AMPK-Mediated Signaling
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AA, compound C, and STO-609 were purchased from Calbiochem (San Diego, CA, USA). Antibodies directed poly(ADP-ribose)polymerase (PARP), caspase-3, B cell lymphoma- (Bcl-) 2, phosphorylated AMPKα, phosphorylated acetyl-CoA carboxylase (ACC), ACC, phosphorylated Ca2+/calmodulin-dependent kinase kinase 2 (CaMKK2), liver kinase B1 (LKB1), and horseradish peroxidase-conjugated secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). Anti-AMPKα and anti-CaMKK2 antibodies were supplied from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-nitrotyrosine (NT) and anti-4-hydroxynonenal (4-HNE) antibodies were purchased from Millipore (Temecula, CA, USA) and Abcam (Cambridge, UK), respectively. Fluo-4-acetoxymethyl ester (Fluo-4) was obtained from Invitrogen (Carlsbad, CA, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), rhodamine 123, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), CCl4, anti-β-actin antibody, and other reagents were supplied from Sigma-Aldrich (St. Louis, MO, USA).
4
Western Blot Analysis of JAK-STAT Signaling
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Protein samples were separated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and transferred onto nitrocellulose membranes (Pall Corp., Port Washington, USA). The membranes were blocked for 1 hours in blocking buffer containing 5% skimmed milk in Tris-buffered saline containing 0.1% Tween 20 and subsequently incubated with the corresponding primary antibodies overnight at 4°C. Blots were washed and incubated with horseradish peroxidase-conjugated secondary antibodies at room temperature for 2 hours, following which the signals were detected using ECL reagent (SurModics, Eden Prairie, USA). Antibodies specific for phospho-JAK1, JAK1, phospho-JAK2, JAK2, phospho-JAK3, JAK3, phospho-tyrosine kinase 2 (TYK2), TYK2, phospho-STAT1, STAT1, phospho-STAT3, poly (ADP-ribose) polymerase (PARP), cleaved caspase 3, B-cell lymphoma (Bcl)-2, Bcl-xL, survivin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were obtained from Cell Signaling Technology (Danvers, USA). Antibodies specific for STAT3, IL-6 receptor α (IL-6Rα), and glycoprotein 130 (gp130) were obtained from Santa Cruz Biotechnology (Santa Cruz, USA), while those for PY-20 and HA were obtained from Abcam (Cambridge, USA).
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