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Protease inhibitor cocktail 5

Manufactured by Merck Group
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The Protease inhibitor cocktail V is a laboratory reagent designed to inhibit the activity of proteases, which are enzymes that break down proteins. This product is commonly used in protein extraction and purification procedures to prevent the degradation of target proteins. The composition and concentration of the inhibitors in the cocktail are optimized to provide broad-spectrum protection against a variety of proteases.

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Lab products found in correlation

Femto chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Mini protean tgx precast gradient gel, by Bio-Rad (2 mentions) Hrp linked goat anti actin, by Santa Cruz Biotechnology (2 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Rabbit anti phospho mtor, by Cell Signaling Technology (1 mentions) Horseradish peroxidase hrp linked anti rabbit igg pab, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Protease Inhibitor Cocktail Set V Protease inhibitor cocktail Proteases inhibitor cocktail Protease cocktails Cocktail inhibitor Protease inhibitors

2 protocols using protease inhibitor cocktail 5

1

Phosphorylation Analysis of mTOR and MERTK

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Cultured cells were lysed in buffer containing NaF, Na4P2O7, Na3VO4, and protease inhibitor cocktail V (EMD Millipore, Temecula, CA) to preserve protein phosphorylation. Proteins were resolved on Mini‐PROTEAN TGX precast gradient gels (BioRad, Berkeley, CA), transferred to polyvinylidene fluoride membranes, and incubated with rabbit anti‐phospho‐mTOR (monoclonal antibody [mAb]; Cell Signaling Technology, Danvers, MA), anti‐mTOR (mAb; Cell Signaling Technology), anti‐phospho‐MERTK (mer receptor tyrosine kinase; polyclonal antibody [pAb]; FabGennix, Frisco, TX), or anti‐MERTK (pAb; FabGennix). The blottings were then incubated with horseradish peroxidase (HRP)‐linked anti‐rabbit immunoglobulin G (pAb; Cell Signaling Technology) and HRP‐linked goat anti‐Actin (Santa Cruz Biotechnology, Santa Cruz, CA) and visualized with an 80/20 mix of SuperSignal West Pico Chemiluminescent Substrate and Femto Chemiluminescent Substrate (Thermo Scientific, Rochester, NY). Densitometry was performed using ImageJ analysis software (National Institutes of Health, Bethesda, MD).
Labonte A.C., Sung S.J., Jennelle L.T., Dandekar A.P, & Hahn Y.S. (2016). Expression of scavenger receptor‐AI promotes alternative activation of murine macrophages to limit hepatic inflammation and fibrosis. Hepatology (Baltimore, Md.), 65(1), 32-43.
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2

Phosphorylation of mTOR and MERTK

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cultured cells were lysed in buffer containing NaF, Na4P2O7, Na3VO4, and protease inhibitor cocktail V (EMD Millipore, Temecula, CA) to preserve protein phosphorylation. Proteins were resolved on Mini-PROTEAN TGX precast gradient gels (BioRad, Berkeley, CA), transferred to PVDF membranes, and incubated with rabbit anti-phospho-mTOR (mAb, Cell Signaling Technology, Danvers, MA), anti-mTOR (mAb, Cell Signaling Technology), anti-phospho-MERTK (pAb, FabGennix, Frisco, TX), or anti-MERTK (pAb, FabGennix). The blots were then incubated with horseradish peroxidase (HRP)-linked anti-rabbit IgG (pAb, Cell Signaling Technology) and HRP-linked goat anti-Actin (Santa Cruz Biotechnology, Santa Cruz, CA) and visualized with an 80/20 mix of SuperSignal West Pico Chemiluminescent Substrate and Femto Chemiluminescent Substrate (Thermo Scientific, Rochester, NY). Densitometry was performed using ImageJ analysis software (National Institute of Health, Bethesda, MD).
Labonte A.C., Sung S.J., Jennelle L.T., Dandekar A.P, & Hahn Y.S. (2016). Expression of scavenger receptor‐AI promotes alternative activation of murine macrophages to limit hepatic inflammation and fibrosis. Hepatology (Baltimore, Md.), 65(1), 32-43.
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