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Protease inhibitor solution

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Protease inhibitor solution is a laboratory product that contains a mixture of chemicals designed to inhibit the activity of proteases, which are enzymes that break down proteins. This solution is commonly used in biochemical and molecular biology applications to preserve the integrity of protein samples during sample preparation and analysis.

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Lab products found in correlation

Tissue homogenizer, by Biospec (3 mentions) 0.45 m filter, by Merck Group (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Protein assay kit, by Bio-Rad (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Carboxyfluorescein diacetate succinimidyl ester, by Thermo Fisher Scientific (1 mentions) Triptolide, by Merck Group (1 mentions) Polyvinylidene difluoride pvdf membrane, by Bio-Rad (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Elisa ready set go, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor cocktails 2 and 3, by Merck Group (1 mentions) Hplc grade acetonitrile, by Thermo Fisher Scientific (1 mentions) Milli q ultrapure water system, by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Methanol, by Thermo Fisher Scientific (1 mentions) Human epidermal growth factor (hegf), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions)

Spelling variants of this product

Protease inhibitors (2755 citations) Protease inhibitor cocktail solution (10 citations) Protease solution (6 citations)

9 protocols using protease inhibitor solution

1

Triptolide Modulates Inflammatory Response

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Triptolide (molecular weight 360.4, purity ≥98% by HPLC), lipopolysaccharide (LPS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO) and protease inhibitor solution were obtained from Sigma Chemical (St. Louis, MO, USA). RPMI-1640 medium, fetal bovine serum (FBS), penicillin, streptomycin, LipofectAMINE 2000 and carboxyfluorescein diacetate succinimidyl ester were purchased from Invitrogen (Camarillo, CA, USA). The cytokine and chemokine ELISA assay kits were obtained from R&D Systems (Minneapolis, MN, USA). RIPA was supplied by Thermo Scientific (Waltham, MA, USA). A protein assay kit and polyvinylidene difluoride (PVDF) membranes were obtained from Bio-Rad Laboratories (Hercules, CA, USA).
Song C., Wang Y., Cui L., Yan F, & Shen S. (2019). Triptolide attenuates lipopolysaccharide-induced inflammatory responses in human endothelial cells: involvement of NF-κB pathway. BMC Complementary and Alternative Medicine, 19, 198.
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2

Testicular Tunica Albuginea Extraction

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The tunica albuginea of the testes was removed. Its content was stored in a protease inhibitor solution (1:250; Sigma, St. Louis, MO, USA) and was homogenized (with the tissues in ice). Lysates were centrifuged (13,000 rpm for 15 min at 4 °C), and the upper phase was filtered through a 0.45 µm filter (Merck KGaA, Darmstadt, Germany) and stored at −80 °C.
Bagdadi N., Sawaied A., AbuMadighem A., Lunenfeld E, & Huleihel M. (2021). The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells. International Journal of Molecular Sciences, 22(3), 1147.
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3

Lung Tissue Homogenization for Cytokine Analysis

Cited 1 time
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After collecting BALF, remaining lung tissue was resected and homogenized using a tissue homogenizer (Biospec Products, Bartlesville, OK, USA) in lysis buffer and protease inhibitor solution (Sigma-Aldrich). After incubation and centrifugation, supernatants were harvested and passed through a 0.45-micron filter (Gelman Science, Ann Arbor, MI, USA). The final preparations were stored at –20°C for cytokine analysis as described previously [19 (link)].
Han H., Oh E.Y., Lee J.H., Park J.W, & Park H.J. (2020). Effects of Particulate Matter 10 Inhalation on Lung Tissue RNA expression in a Murine Model. Tuberculosis and Respiratory Diseases, 84(1), 55-66.
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4

Testicular Protein Extraction and Preservation

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The tunica albuginea were removed. Then, the testes were stored in a protease inhibitor solution (1:250; Sigma, St. Louis, MN, USA) at −20 °C. Thereafter, the tissues were homogenized (with the tissues in ice). The homogenates were centrifuged (13,000 rpm for 15 min at 4 °C), and the upper phase was collected and filtered through a 0.45 µm filter (Merck KGaA, Darmstadt, Germany) and stored at −80 °C.
AbuMadighem A., Solomon R., Stepanovsky A., Kapelushnik J., Shi Q., Meese E., Lunenfeld E, & Huleihel M. (2018). Development of Spermatogenesis In Vitro in Three-Dimensional Culture from Spermatogonial Cells of Busulfan-Treated Immature Mice. International Journal of Molecular Sciences, 19(12), 3804.
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5

Cytokine Quantification in Lung Homogenates

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After collecting BAL fluid, the right lung was homogenized using tissue homogenizer (Biospec Products, Bartlesville, USA) in 1.5 ml of RIPA buffer (Thermo, IL, USA) and protease inhibitor solution (Sigma-aldrich). After incubation for 30 min on ice, homogenates were centrifuged at 14,000 x g for 20 min. Supernatants were collected. Concentrations of interleukin-5 (IL-5) (detection limit: 31.2 pg/ml), interleukin-13 (IL-13) (detection limit: 62.5 pg/ml), interleukin-1beta (IL-1β) (detection limit: 15.6 pg/ml), and interferon gamma (IFN-γ) (detection limit: 31.2 pg/ml) in lung homogenate were measured by ELISA (R&D Systems, San Diego, USA) according to the manufacturer’s instructions.
Han H., Park Y.H., Park H.J., Lee K., Um K., Park J.W, & Lee J.H. (2016). Toxic and adjuvant effects of silica nanoparticles on ovalbumin-induced allergic airway inflammation in mice. Respiratory Research, 17, 60.
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6

Quantification of Colonic Cytokines

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A 50 mM Tris-HCl with 10 μg/mL protease inhibitor solution (Sigma-Aldrich Co., USA) was used to homogenise the colonic samples over ice. Centrifugation of 20 minutes was used on the homogenates at 30,000 ×g (4°C). Following centrifugation, a Sandwich ELISA Kit (ELISA Ready-SET-GO, eBioscience, CA, USA) was used to test the supernatants for TNF-α and IL-6. Normalization of cytokine was carried out to match the colonic samples' protein levels.
Guan G., Wang H., Chen S., Liu G., Xiong X., Tan B., Duraipandiyan V., Al-Dhabi N.A, & Fang J. (2016). Dietary Chitosan Supplementation Increases Microbial Diversity and Attenuates the Severity of Citrobacter rodentium Infection in Mice. Mediators of Inflammation, 2016, 9236196.
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7

Proteomics Analysis of SKBR3 Breast Cancer

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SKBR3 breast cancer cells, phosphate buffered saline (PBS) and trypsin/EDTA were purchased from the American Tissue Culture Collection (ATCC, Manassas, VA, USA). Fetal bovine serum (FBS) was from Gemini-Bio Products (West Sacramento, CA, USA), McCoy’s 5A cell culture medium from Life Technologies (Carlsbad, CA, USA), human epidermal growth factor (hEGF) from PeproTech (Rocky Hill, NJ), Normocin from InvivoGen (San Diego, CA, USA), and sequencing grade modified trypsin and Lys-C from Promega Corporation (Madison, WI, USA). Urea, dithiothreitol (DTT), acetic acid, trifluoroacetic acid (TFA), ammonium bicarbonate, sodium chloride, Trizma base and hydrochloride buffer, protease inhibitor solution and phosphatase inhibitor cocktails 2 and 3 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Zorbax SB-C18/5 μm particles, SPEC-PTC18 and SPEC-PTSCX solid-phase extraction pipette tips were purchased from Agilent Technologies (Santa Clara, CA, USA), and fused silica capillaries from Polymicro Technologies. HPLC-grade acetonitrile and methanol were from Fisher Scientific (Fair Lawn, NJ, USA), and DI water was prepared with a MilliQ Ultrapure water system (Millipore, Bedford, MA, USA).
Deng J., Julian M.H, & Lazar I.M. (2018). Partial Enzymatic Reactions: A Missed Opportunity in Proteomics Research. Rapid communications in mass spectrometry : RCM, 32(23), 2065-2073.
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8

Lung Tissue Homogenization and Cytokine Analysis

Cited 1 time
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After collecting BALF, remaining lung tissue was resected and homogenized using a tissue homogenizer (Biospec Products, Bartlesville, OK, USA) in lysis buffer and protease inhibitor solution (Sigma-Aldrich, St Louis, MO, USA). After incubation and centrifugation, supernatants were harvested and passed through a 0.45-micron filter (Gelman Science, Ann Arbor, MI, USA). The final preparations were stored at − 20℃ for cytokine analysis as described previously [17 (link)].
Choi Y.J., Han H., Lee J.H., Lee J., Kim C.Y., Byun M.K., Cho J.H, & Park H.J. (2023). Particulate matter10-induced airway inflammation and fibrosis can be regulated by chitinase-1 suppression. Respiratory Research, 24, 85.
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9

Cytokine Extraction from Ileal Mucosa

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To extract cytokines from the ileal mucosa, 1 ml of lysis buffer composed of RIPA buffer with 1 % protease inhibitor solution (Sigma) was added to 100 mg of scraped mucosa (Leica), and mixed three times for 15 s at 6000 rpm. After centrifugation at 10 000 g for 15 min at 4°C, the supernatant was collected and stored at -80°C. Concentrations of IL-4, IFN-γ, TNF-α and IL-6 were measured using capture sandwich ELISA (porcine Duoset ® ELISA kit; R&D Systems) according to the manufacturer's instructions.
Le Bourgot C., Le Normand L., Formal M., Respondek F., Blat S., Apper E., Ferret-Bernard S, & Le Huërou-Luron I. (2017). Maternal short-chain fructo-oligosaccharide supplementation increases intestinal cytokine secretion, goblet cell number, butyrate concentration and Lawsonia intracellularis humoral vaccine response in weaned pigs. The British journal of nutrition, 117(1).
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