The MB49 cells and MCSCs were respectively harvested. Equal amount proteins were extracted from cells, and separated by 10% sodium dodecyl sulfate -polyacrylamide gel electrophoresis followed by transferring to polyvinylidene difluoride membranes (Millipore, Billerica, MA). The membranes were blocked with 5% skim milk in PBS, and incubated overnight at 4 °C with the primary antibodies including anti-S100A4 (Abcam, Cambridge, MA), anti- IKK (Abcam) and anti-β-actin antibody (Abcam) followed by secondary antibodies (Abcam). Bands were visualized using Fluor Chem FC2 (Alpha Innotech, San Leandro, CA).
Anti ikk
Manufactured by Abcam
Anti-IKK is a primary antibody that recognizes the inhibitor of nuclear factor kappa-B kinase subunit beta (IKK-beta) protein. IKK-beta is a serine/threonine protein kinase that is a key component of the NF-kB signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Anti s100a4, by Abcam (1 mentions)
Fluorchem fc2, by Bio-Techne (1 mentions)
Anti β actin antibody, by Abcam (1 mentions)
Enhanced chemiluminescence system, by GE Healthcare (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Anti gapdh, by Proteintech (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
2 protocols using anti ikk
1
Protein Expression in MB49 and MCSCs
2
Western Blotting Analysis of IKK
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After transfection, cells were lysed with RIPA buffer (Beyotime, Haimen, China) and complete protease inhibitor cocktail (Biotool, Houston, TX). After ultrasonic pyrolysis, the protein concentration was determined with a BCA Protein Assay Kit (Beyotime). The protein samples were then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis electrophoresis. The membrane was blocked in 5% fat-free milk for 2 h, incubated in the primary antibody (anti-IKK, Abcam, Cambridge, MA; anti-GAPDH, Proteintech, Chicago, IL), and washed three times for 5 min each with PBS. After washing, the secondary goat anti-mouse IgG antibody (Jackson ImmunoResearch Laboratories, West Grove, PA) was used at room temperature for 2 h. The enhanced chemiluminescence system (GE Healthcare, Chicago, IL) was applied to detect the results.
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