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Protease inhibitor cocktail p840

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Protease Inhibitor Cocktail P840 is a laboratory reagent designed to inhibit a broad range of protease enzymes. It is commonly used in protein extraction and purification protocols to prevent proteolytic degradation of target proteins.

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Lab products found in correlation

Hrp conjugated goat anti rabbit secondary antibody, by Santa Cruz Biotechnology (3 mentions) Tris glycine sds buffer, by Bio-Rad (3 mentions) Anti ace 2 polyclonal antibody, by Abcam (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Imr 90 cell line, by American Type Culture Collection (1 mentions) α sma, by Cell Signaling Technology (1 mentions) β catenin, by Cell Signaling Technology (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Antibody against β actin, by Cell Signaling Technology (1 mentions)

3 protocols using protease inhibitor cocktail p840

1

Western Blot Analysis of ACE-2, Caspase 9, and ADAM17

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Cells were harvested with protease inhibitor (Protease Inhibitor Cocktail P840, Sigma, St Louis, MO). Soluble protein lysate (45 µg) were loaded and run on 10% Tris.HCL polyacrylamide gels, separated by SDS-PAGE, in 10× Tris/glycine/SDS buffer (BioRad, Berkeley, CA). Gels were transferred to Immun-blot polyvinylidene fluoride blotting membrane. Blotting membrane was blocked by 5% nonfat dry milk in 0.1% Tween 20 in Tris-buffered saline. Western blot analysis of ACE-2 was performed with anti-ACE-2 polyclonal antibody (1:4,000: Abcam Biotechnology, Cambridge, MA). Antibodies for caspase 9 and ADAM17/TACE were obtained from Cell Signaling, Danvers, MA. To ensure equal loading of proteins, membranes were probed with an antibody against β-actin (Cell Signaling Technology). Bands were visualized by HRP-conjugated goat anti-rabbit secondary antibody (1:10,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA) using enhanced chemiluminescence detection by standard techniques.
Oarhe C.I., Dang V., Dang M., Nguyen H., Gopallawa I., Gewolb I.H, & Uhal B.D. (2015). Hyperoxia downregulates angiotensin-converting enzyme-2 in human fetal lung fibroblasts. Pediatric Research, 77(5), 656-662.
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2

Fetal Lung Fibroblast Hypoxia/Hyperoxia Protocol

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The fetal lung fibroblast IMR 90 cell line was purchased from ATCC (Manassas, VA). 6-well collagen I coated plates were obtained from BD Biosciences (Bedford, MA). ACE-2 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). αSMA, NCadherin, β-catenin and β-actin antibodies were purchased from Cell Signaling Technology (Danvers, MA). Oxygen delivery was confirmed in the hypoxia/hyperoxia chamber outlet using a Maxtec MaxO2+A Oxygen Analyzer - R217P62 (Salt Lake City, Utah). Protease Inhibitor Cocktail P840 was purchased from Sigma (Saint Louis, MO) and the Pierce BCA Protein Assay Kit from Thermo Fisher Scientific Pierce Biotechnology (Rockford, IL), 10X Tris/glycine/SDS buffer was obtained from BioRad (Berkeley, CA), and HRP-conjugated goat anti-rabbit secondary antibody from Santa Cruz Biotechnology (Santa Cruz, CA). All other materials were of reagent grade.
Mohamed T.L., Nguyen H.T., Abdul-Hafez A., Dang V.X., Dang M.T., Gewolb I.H, & Uhal B.D. (2016). PRIOR HYPOXIA PREVENTS DOWNREGULATION OF ACE-2 BY HYPEROXIA IN FETAL HUMAN LUNG FIBROBLASTS. Experimental lung research, 42(3), 121-130.
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3

Western Blot Analysis of ACE-2, Caspase 9, ADAM17

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were harvested with protease inhibitor (Protease Inhibitor Cocktail P840, Sigma, St Louis, MO). Soluble protein lysate (45 μg) were loaded and run on 10% Tris.HCL polyacrylamide gels, separated by SDS-PAGE, in 10× Tris/glycine/SDS buffer (BioRad, Berkeley, CA). Gels were transferred to Immun-blot PVDF blotting membrane. Blotting membrane was blocked by 5% nonfat dry milk in 0.1% Tween 20 in Tris-buffered saline. Western blot analysis of ACE-2 was performed with anti-ACE-2 polyclonal antibody (1:4,000: Abcam Biotechnology, Cambridge, MA). Antibodies for caspase 9 and ADAM17/TACE were obtained from Cell Signaling, Danvers, MA. To ensure equal loading of proteins, membranes were probed with an antibody against β-actin (Cell Signaling Technology). Bands were visualized by HRP-conjugated goat anti-rabbit secondary antibody (1:10,000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA) using enhanced chemiluminescence detection by standard techniques.
Oarhe C.I., Dang V., Dang M., Nguyen H., Gopallawa I., Gewolb I.H, & Uhal B.D. (2015). Hyperoxia downregulates angiotensin-converting enzyme-2 in human fetal lung fibroblasts. Pediatric Research, 77(5), 656-662.
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