T47D cells were plated as on 24-well plates at 70–87% confluency and allowed to adhere overnight at 37 °C. Following overnight incubation, cells were serum starved with serum free RPMI media for 4 h and subsequently treated with 25 ng/mL chOSM (PeproTech, Rocky Hill, NJ; Cat# 300-10 T) or rhOSM at 25 ng/mL for 30 min. Afterwards, lysates were collected as previously described for immunoblot20 (link). Lysate samples were diluted 1:4 in 1X PBS supplemented with 1% BSA. Levels of phospho-STAT3 were detected using the PathScan Phospho-STAT3 (Y705) Sandwich ELISA kit (Cell Signaling Technologies; Cat# 7300) as per manufacturer's instructions. Relative expression was quantified comparing non-treated and rhOSM treated samples to chOSM levels. Experiments were run in triplicate and data was analyzed using Microsoft Excel and Prism 8 (v. 8.2.1, GraphPad Software).
Pathscan phospho stat3 y705 sandwich elisa kit
Manufactured by Cell Signaling Technology
The PathScan Phospho-STAT3 (Y705) Sandwich ELISA kit is a quantitative immunoassay designed to detect and measure the level of phosphorylated STAT3 (Tyr705) in cell lysates. The kit uses a capture antibody specific for STAT3 and a detection antibody specific for the phosphorylated form of STAT3 at Tyr705.
Automatically generated - may contain errors
Lab products found in correlation
Chosm, by Thermo Fisher Scientific (1 mentions)
Rhosm, by Thermo Fisher Scientific (1 mentions)
Prism 8, by GraphPad (1 mentions)
Complete protease inhibitor and phosphatase inhibitor cocktail tablets, by Roche (1 mentions)
Protein assay kit, by Bio-Rad (1 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
2 protocols using pathscan phospho stat3 y705 sandwich elisa kit
1
Comparative Analysis of chOSM and rhOSM
2
Quantitative STAT3 Phosphorylation Assay
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