Colonic tissues were lysed in Cell Lytic Mammalian Tissue Lysis Extraction Reagent (Sigma) and analyzed using the 25-analyte MILLIPLEX MAP Mouse Cytokine/Chemokine Magnetic Bead Panel (EMD Millipore) on a FLEXMAP 3D instrument (Luminex) (21 (link), 22 (link)). Data were standardized to tissue protein concentrations measured by the BCA Protein Assay Kit (Pierce). TNF-α and IFN-γ in tissues was measured using R&D DuoSet ELISA kits (10 (link)).
Cell lytic mammalian tissue lysis extraction reagent
Manufactured by Merck Group
The Cell Lytic Mammalian Tissue Lysis Extraction Reagent is a solution designed for the extraction and lysis of proteins from mammalian cell and tissue samples. The reagent facilitates the release of cellular components, including proteins, for further analysis or processing.
Automatically generated - may contain errors
4 protocols using cell lytic mammalian tissue lysis extraction reagent
1
Cytokine and Chemokine Profiling in Colonic Tissues
2
Multiplex Cytokine Profiling of Colon Tissue
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Colon tissues were lysed in Cell Lytic Mammalian Tissue Lysis Extraction Reagent (Sigma) and analyzed using the 32-analyte MILLIPLEX MAP Mouse Cytokine/Chemokine Magnetic Bead Panel (Millipore Sigma) on a FLEXMAP 3D instrument (Luminex) as reported (Coburn et al., 2018 (link); Singh et al., 2018 (link)). Data were standardized to tissue protein concentrations measured by the BCA Protein Assay Kit (Pierce).
3
Cytokine and Chemokine Profiling in Colonic Tissues
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Colonic tissues were lysed in Cell Lytic Mammalian Tissue Lysis Extraction Reagent (Sigma) and analyzed using the 25-analyte MILLIPLEX MAP Mouse Cytokine/Chemokine Magnetic Bead Panel (EMD Millipore) on a FLEXMAP 3D instrument (Luminex) [9 (link), 10 (link)]. Data were standardized to tissue protein concentrations measured by the BCA Protein Assay Kit (Pierce).
4
Colonic Protein Expression Analysis
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Colonic tissues or cells were lysed in Cell Lytic Mammalian Tissue Lysis Extraction Reagent (Sigma) containing the EDTA-free Protease Inhibitor Cocktail Set III, and the Phosphatase Inhibitor Set I (Millipore), and disrupted by sonication at 40 W. Protein concentrations were determined by the BCA Protein Assay Kit (Pierce). Total proteins (80 μg per lane) were separated on 10% polyacrylamide gels (Bio-Rad) and transferred onto PVDF membranes. Monoclonal anti-EspB (1:1000), rabbit monoclonal anti-Talin-1 Ab (1:2000), rabbit polyclonal anti-ACTN1 Ab (1:4000; Cell Signaling), rabbit polyclonal anti-SLC7A2 Ab (1:2000; Abcam), and mouse monoclonal anti-β-actin (1:10,000; Sigma) were used as described [9 (link), 10 (link)].
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