Protein expression was determined by Western blotting, which was performed according to the method of Lee et al. [49 (link)]. Briefly, proteins from cellular lysates were separated on SDS-PAGE gels and transferred to PVDF membranes (Millipore, Danvers, MA, USA). After blocking for 1 h with 5% skim milk in TBST buffer, the membranes were incubated overnight at 4 °C with antibodies specific for the following proteins: iNOS (BD Biosciences, San Jose, CA, USA), COX-2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), CD80, CD86, TNF-α, phosphor-PI3K p85 (Tyr458), PI3K, phosphor-AKT (T308), AKT, phospho-p70S6K (T389), p70S6K, phosphor-NF-κB (Ser536), NF-κB, PDK1, LDHA (Cell Signaling Technology; Danvers, MA, USA), HIF1-α (Novus Biologicals, Building IV Centennial, CO, USA), and β-actin (Sigma). After three washes in TBST, membranes were incubated with HRP-conjugated secondary antibodies (GeneTex, Irvine, CA, USA). HRP was detected using the ECL reagent (BioNote, Hwaseong, Gyeonggi-do, Korea).
Phosphor pi3k p85
Manufactured by Cell Signaling Technology
Sourced in United States
The Phosphor-PI3K p85 is a laboratory equipment product that is used to detect and measure the phosphorylation of the p85 subunit of phosphoinositide 3-kinase (PI3K). This product is designed to provide researchers with a reliable and accurate tool for studying the activation of the PI3K signaling pathway, which is involved in various cellular processes such as cell growth, survival, and metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (1 mentions)
Hrp conjugated secondary antibody, by GeneTex (1 mentions)
Phosphor akt, by Cell Signaling Technology (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Tnf α, by Cell Signaling Technology (1 mentions)
Phosphor nf κb, by Cell Signaling Technology (1 mentions)
Cox 2, by Santa Cruz Biotechnology (1 mentions)
Hif 1α, by Novus Biologicals (1 mentions)
Phospho p70s6k, by Cell Signaling Technology (1 mentions)
Nf κb, by Cell Signaling Technology (1 mentions)
Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions)
S6 ribosomal protein, by Cell Signaling Technology (1 mentions)
Aim 5 medium, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Antibodies to phosphoakt, by Cell Signaling Technology (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Mcl 1, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
2 protocols using phosphor pi3k p85
1
Protein Expression Analysis by Western Blotting
2
Western Blot Antibody Validation Protocol
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