The reagents used in this study were obtained from the indicated suppliers: IFN-γ, IL-1β, TNF-α, IL-4, IL-13, and TGF-β from R&D Systems (Minneapolis, MN, USA); antibodies against α-SMA (ab7817) from Abcam (Cambridge, UK); antibodies against GAPDH (sc47724) and TSG-6 (sc377277) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against STAT1 (9172S), pSTAT1 (9167S), STAT3 (9139S), pSTAT3 (9145S), and Smad2/3 (8685S) from Cell Signaling Technology (Danvers, MA, USA); antibodies against pSmad2/3 (PA5-110155) from ThermoFisher Scientific (Waltham, MA, USA); chemical inhibitors for JAK1 (Abrocitinib, HY-107429; MedChemExpress, Princeton, NJ, USA) and JAK2 (AG-490, S1143; Selleck Chemicals, Houston, TX, USA); scramble and TSG-6 siRNA (Santa Cruz). All other materials were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise indicated.
Pa5 110155
Manufactured by Thermo Fisher Scientific
Sourced in United States, Australia
The PA5-110155 is a laboratory equipment product from Thermo Fisher Scientific. It is designed to perform a specific core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Smad2 3 8685, by Cell Signaling Technology (1 mentions)
Ag490 s1143, by Selleck Chemicals (1 mentions)
Antibodies against α sma ab7817, by Abcam (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by R&D Systems (1 mentions)
Tgf β, by R&D Systems (1 mentions)
Il 13, by R&D Systems (1 mentions)
Ab27969, by Abcam (1 mentions)
S2369, by Agilent Technologies (1 mentions)
Decloaking chamber, by Biocare Medical (1 mentions)
K5007, by Agilent Technologies (1 mentions)
Ab32572, by Abcam (1 mentions)
2 protocols using pa5 110155
1
Modulation of Inflammatory Pathways
2
Immunohistochemical Analysis of EndMT Drivers
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The immunohistochemistry method was used for all EndMT drivers. Briefly, lung tissue sections were deparaffinised in xylene and ethanol, respectively, followed by antigen retrieval performed at 110 °C for 15 min using target retrieval citrate buffer pH 6.0 (Dako S2369, Mulgrave, VIC, Australia) in a decloaking chamber (Biocare Medical, Pacheco, CA, USA). Primary antibodies were used to stain tissues for 60 min: mouse monoclonal TGF-β1 (1:2000, Abcam ab27969, Melbourne, VIC, Australia), polyclonal phospho-Smad2/Smad3 (pSmad-2/3, 1:100, ThermoFisher PA5-110155, Scoresby, VIC, Australia), mouse monoclonal Smad-7 (1:50, Santa Cruz Biotechnology sc-101152, Dallas, TX, USA), and rabbit monoclonal anti-β-catenin (1:100, Abcam ab32572, Melbourne, VIC, Australia), followed by secondary HRP rabbit/mouse antibodies (Dako K5007, Mulgrave, VIC, Australia). The DAB substrate (Dako K5007, Mulgrave, VIC, Australia) was added to make the protein markers visible, and hematoxylin (Australian Biostain P/L, Traralgon, Australia) was used to counterstain the nucleus. We have previously published using these techniques [4 (link),11 (link),12 (link),13 (link),14 (link),15 (link),16 (link),17 (link)].
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