Anti-ALK, anti-phospho-Akt at serine (Ser) 473 (pAkt), anti-Akt, anti-Slug, anti-Snail, and anti-cleaved caspase 3 antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-Sox11 and anti-β-actin antibodies and doxorubicin were obtained from Sigma-Aldrich Chemicals (St. Louis, MO, USA). Anti-N-myc, anti-Twist1, and anti-Histone H1 antibodies were from Abcam (Cambridge, MA, USA). Anti-NF-κB/p65, anti-p27kip1, and anti-bax antibodies were from BD Biosciences (San Jose, CA, USA). Anti-bcl-2 and anti-p21waf1 antibodies were from Dako (Glostrup, Denmark). Anti-cyclin A antibody was from Novocastra (Newcastle, UK). Recombinant human tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and hepatocyte growth factor (HGF) were purchased from R&D Systems (Minneapolis, MN, USA).
Anti n myc
Manufactured by Abcam
Sourced in United States
Anti-N-myc is a lab equipment product that targets the N-myc protein. It is used for research purposes in cellular and molecular biology studies.
Automatically generated - may contain errors
Lab products found in correlation
Anti cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Doxorubicin, by Merck Group (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti histone h1, by Abcam (1 mentions)
Transforming growth factor β1, by R&D Systems (1 mentions)
Anti p27kip1, by BD (1 mentions)
Anti alk, by Cell Signaling Technology (1 mentions)
Anti bax, by BD (1 mentions)
Recombinant human tumor necrosis factor α, by R&D Systems (1 mentions)
Anti snail, by Cell Signaling Technology (1 mentions)
Anti slug, by Cell Signaling Technology (1 mentions)
Anti twist1, by Abcam (1 mentions)
Anti nf κb p65, by BD (1 mentions)
Horseradish peroxidase labeled secondary antibody, by Agilent Technologies (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Image lab software 5, by Bio-Rad (1 mentions)
Anti vegfa, by Abcam (1 mentions)
Lumi light pod substrate, by Roche (1 mentions)
Anti bcl2, by Agilent Technologies (1 mentions)
4 protocols using anti n myc
1
Antibody Characterization for Cell Signaling
2
Western Blot Analysis of Protein Expression
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Whole-cell lysates for western blot analysis were prepared using standard radioimmunoprecipitation assay buffer. In brief, 30 μg protein were separated on 10% acrylamide/bis-acrylamide gels before transfer to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA, USA) using CAPS buffer (10 mM 3-[cyclohexylamino]-1-propane sulfonic acid, pH 11; 10% methanol). The following primary antibodies were diluted in 5% w/v BSA (Sigma) in TBS-T: 1:1000 anti-Bcl-2 (Dako), 1:1000 anti-cleaved caspase 3 (Cell Signaling Technologies, Danvers, MA, USA), 1:30 000 anti-tubulin (Promega, Madison, WI, USA), 1:1000 anti-VEGFA (vascular endothelial growth factor, Abcam) and 1:1000 anti-NMYC (Abcam). Horseradish peroxidase-labeled secondary antibodies were used (Dako) and detection was carried out with Lumi-Light POD-substrate (Roche, Basel, Switzerland). Quantification of band intensities was performed using Image Lab Software 5.2.1 (Bio-Rad) and adapted to the corresponding loading control.
3
Subcellular Fractionation and Phospho-protein Analysis
4
EV Protein Characterization by Western Blot
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EVs were lysed with RIPA buffer (Abcam, Inc., Cambridge, UK) containing 60 mg/mL dithiothreitol and SIGMAFAST™ Protease Inhibitor tablets (Sigma-Aldrich, St. Louis, MO, USA) and the protein amounts were quantitated using a Bradford assay. Proteins (20 µg) were separated by 12.5% SDS-PAGE and transferred onto a polyvinylidene difluoride (PVDF) membrane (Millipore Immobilon®-P PVDF Membrane, Merck KGaA, Darmstadt, Germany). After blocking with 5% skim milk, the membrane was incubated with a 1:1000 primary antibody as follows: anti-Alix (#2171; Cell Signaling, Danvers, MA, USA), anti-Hsp70 (ab79852; Abcam plc, Cambridge, UK), anti-Tsg101 (ab125011; Abcam plc.), anti-n-Myc (ab24193; Abcam plc.), or anti-calnexin (#40090; Cell Signaling), at 4 °C overnight. The probed membrane was washed and incubated with 1:1000 anti-rabbit (P0448; Dako, Glostrup, Denmark) or 1:2000 anti-mouse (P0447; Dako) as an appropriate at room temperature for 1 h. The detected protein bands were enhanced by SuperSignal West Pico chemiluminescence substrate (Thermo Scientific) and captured by G:BOX Chemi XRQ (SYNGENE, Cambridge, CB4 1TF, UK).
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