Millipore
Rabbit polyclonal anti-NOS2/iNOS antibody
AB5382
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In summary, Western blots showed that tumors derived from iNOS shRNA-targeted LCSCs exhibited less TACE and NICD than scrambled shRNA-treated cells.
Western Blot
Mouse
1/5000
5% milk
1/2000
Room temperature 1 hour
ECL
Cells were lysed with the buffer [1% SDS, 10 mmol/L Tris-Cl (pH 7.6), 20 µg/µL aprotinin, 20 µg/µL leupeptin, and 1 mmol/L 4-(2-aminoethyl)benzenosulfonyl fluoride]. The protein concentrations were determined using the Bicinchoninic Acid Protein Assay kit (Pierce, Rockford, IL). Twenty micrograms of protein were separated on SDS-PAGE gels and transferred to polyvinylidene difluoride membranes. After blocking, the membranes were incubated with the appropriate primary antibody at 4°C overnight. After washing with TBST, the membranes were incubated with HRP-conjugated secondary antibodies at room temperature for 1 hour. Proteins were detected with the enhanced chemiluminescence kit (Amersham Pharmacia Biotechnology, Inc., Piscataway, NJ). Western blots showed that tumors derived from iNOS shRNA-targeted LCSCs exhibited less TACE and NICD than scrambled shRNA-treated cells.
doi/10.1073/pnas.1722100115
Figure 2C
Worked well
N/A
Worked as expected