The tissue proteins were separated by electrophoresis on sodium dodecyl sulfate- (SDS-) polyacrylamide gels and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Temecula, CA) using a semidry transfer system. The membranes were first incubated in blocking solution (5% skim milk) and then incubated overnight at 4°C with the primary antibodies of iNOS, COX2, IκBα, p-IκBα, p65, p-p65, and β-actin (Sigma, St. Louis, MO, USA). The β-actin was used as internal control. Immunoblots were incubated with the corresponding secondary antibody conjugated with horseradish peroxidase for 1 h. Membranes were developed using an electrochemiluminescence kit (Merck, China). The density of the immunoreactive bands was analyzed using Image J software (National Institutes of Health, Bethesda, MD, USA).
P iκbα
Manufactured by Merck Group
Sourced in United States
P-IκBα is a lab equipment product that serves as a key regulator of the NF-κB signaling pathway. It functions to sequester NF-κB transcription factors in the cytoplasm, thereby inhibiting their nuclear translocation and transcriptional activity.
Automatically generated - may contain errors
Lab products found in correlation
Electrochemiluminescence kit, by Merck Group (1 mentions)
Cox 2, by Merck Group (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Cell lysis buffer 10, by Cell Signaling Technology (1 mentions)
Rt pcr kit, by Toyobo (1 mentions)
Myd88, by Merck Group (1 mentions)
Traf6, by Merck Group (1 mentions)
Nf κb p65, by Merck Group (1 mentions)
Keap1, by Merck Group (1 mentions)
Gapdh, by Merck Group (1 mentions)
Trypsin, by Merck Group (1 mentions)
2 7 dichlorofluorescein diacetate dcf da, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Roche (1 mentions)
Catalase, by Merck Group (1 mentions)
Superoxide dismutase, by Merck Group (1 mentions)
E cadherin, by Merck Group (1 mentions)
N cadherin, by Merck Group (1 mentions)
P erk1 2, by Merck Group (1 mentions)
Mmp 9, by Merck Group (1 mentions)
3 protocols using p iκbα
1
Western Blot Analysis of Inflammatory Proteins
2
Molecular Mechanisms of Oxidative Stress
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Antibodies against KDR, CD29, CD90, CD44, CD105, CD34, Von Willenbrand factor (vWF), GAPDH, NOX1, NOX4, HO-1, TLR4, MyD88, TRAF6, NFκB P65, p-IκBα, Keap-1, Nrf2 and 2′,7′-dichlorofluorescein diacetate (DCF-DA) were obtained from Sigma (St Louis, MO, USA). DMEM (high glucose), serum-free EBM, and fetal bovine serum (FBS) were from Nego (Shanghai, China). Cell lysis buffer (10 ×) was obtained from Cell Signaling Technology (CST, Boston, USA). The RT-PCR kit was purchased from TOYOBO (Shanghai, China). Other reagents included DAPI (Roche, Basel, Germany), hematoxylin and eosin (H&E; Toronto Chemicals, Toronto, ON, Canada), and trypsin (Sigma). All pairs of real-time PCR primers were synthesized by Shenggong Biotechnology (Shanghai, China). Other chemicals and reagents were of analytical grade.
3
Comprehensive Molecular Profiling of Cell Culture
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McCoy's 5A and F-12 K media and other reagents for cell culture were purchased from Carlo Erba Reagents (Milan, Italy). The EvaGreen 2X qPCRMaster Mix kit and the primers (Supplementary Table 1) for real time PCR were obtained from Applied Biological Materials Inc. (Canada) and Sigma-Aldrich (Milan, Italy). The primary and secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA) and Sigma-Aldrich (Milan, Italy).
The following antibodies were used: Nrf2, superoxide dismutase (SOD), catalase, heme oxygenase 1 (HO-1), NF-κB, phosphorylated inhibitor of kappa B (p-IκBα), p53, caspase-3, cleaved-poly (ADP-ribose) polymerase (c-PARP), EGFR, human epidermal growth factor receptor 2 (HER2), p-Akt, p-mammalian target of rapamycin (p-mTOR), p-p38MAPK, p-extracellular-signal-regulated kinase 1/2 (p-Erk1/2), matrix metalloproteinases 2 (MMP-2), MMP-9, E-cadherin, N-cadherin, β-catenin and glyceraldehyde-3-phosphate dehydrogenase (GADPH). All other chemicals were purchased from Sigma-Aldrich (Milan, Italy). MH samples inventing from New Zealand were imported to Italy by EfitSrl and kept at 4°C until analysis.
The following antibodies were used: Nrf2, superoxide dismutase (SOD), catalase, heme oxygenase 1 (HO-1), NF-κB, phosphorylated inhibitor of kappa B (p-IκBα), p53, caspase-3, cleaved-poly (ADP-ribose) polymerase (c-PARP), EGFR, human epidermal growth factor receptor 2 (HER2), p-Akt, p-mammalian target of rapamycin (p-mTOR), p-p38MAPK, p-extracellular-signal-regulated kinase 1/2 (p-Erk1/2), matrix metalloproteinases 2 (MMP-2), MMP-9, E-cadherin, N-cadherin, β-catenin and glyceraldehyde-3-phosphate dehydrogenase (GADPH). All other chemicals were purchased from Sigma-Aldrich (Milan, Italy). MH samples inventing from New Zealand were imported to Italy by EfitSrl and kept at 4°C until analysis.
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