Tumor (non‐necrotic) and liver tissues were homogenized in radio immunoprecipitation assay buffer (150 mM NaCl, 1% NP‐40, 0.5% Na‐deoxycholate, 0.1% sodium dodecyl sulfate, and 50 mM Tris‐HCl pH 7.4) containing protease and phosphatase inhibitors (Roche, Rotkreuz, Switzerland). Protein concentration was measured using the Pierce BCA Protein Assay (Thermo Fisher Scientific, Rockford, IL). Immunoblots were performed with antibodies against protein kinase B (AKT; Ser473), signal transducer and activator of transcription 3 (STAT3), phospho‐STAT3, AMPKα, phospho‐AMPKα (Thr172), raptor, phospho‐raptor (Ser792), S6‐ribosomal protein (S6‐RP), phospho‐S6‐RP (Ser240/244), fatty acid synthase (FAS), acetyl‐coenzyme A (CoA) carboxylase (ACC), phospho‐ACC (Ser79), 4E‐binding protein 1, phospho‐4E‐binding protein 1, mTOR, vinculin, and PTEN (Cell Signaling). Signals were analyzed using the Bio‐1D Advanced software (Vilber‐Lourmat, Marne‐la‐Vallée, France).
4e binding protein 1
Manufactured by Cell Signaling Technology
4E-binding protein 1 (4E-BP1) is a key regulator of protein synthesis and cell growth. It functions by binding to and inhibiting the eukaryotic translation initiation factor 4E (eIF4E), which is essential for the initiation of protein translation. This interaction prevents the formation of the eIF4F complex, thereby suppressing protein synthesis.
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Lab products found in correlation
Protease and phosphatase inhibitor, by Roche (1 mentions)
Bio 1d advanced software, by Vilber (1 mentions)
Vinculin, by Cell Signaling Technology (1 mentions)
Pierce bca protein assay, by Thermo Fisher Scientific (1 mentions)
Phospho p70 s6 kinase t389, by Cell Signaling Technology (1 mentions)
Antirabbit 111 035 144, by Jackson ImmunoResearch (1 mentions)
P70 s6 kinase, by Cell Signaling Technology (1 mentions)
2 protocols using 4e binding protein 1
1
Liver and Tumor Protein Signaling Analysis
2
Immunoblotting Analysis of Cellular Proteins
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For immunoblotting, the following antibodies were used: a mouse monoclonal antibody against heat shock protein 90 kDa (610419; BD Transduction Lab) and rabbit polyclonal antibodies against PFAS (A17517; Abclonal), 4E-binding protein 1 (9452; Cell Signaling Technology), p70 S6 Kinase (9202; Cell Signaling Technology), phospho-p70 S6 Kinase (T389) (9234; Cell Signaling Technology), LC3 (63 (link)), TSC2 (3990; Cell Signaling Technology), CAD (11933; Cell Signaling Technology), and DHODH (14877-1-AP; Proteintech). For secondary antibodies, horseradish peroxidase–conjugated anti-mouse (111-035-003; Jackson ImmunoResearch Laboratories, Inc) and anti-rabbit (111-035-144; Jackson ImmunoResearch Laboratories, Inc) IgGs were used.
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