Protease inhibitor

Manufactured by Roche

Sourced in United States, Switzerland, Germany, China, United Kingdom, Canada, France, Japan, Italy, Australia, Spain, India, Macao, Netherlands, Denmark, Belgium, Cameroon

Protease inhibitors are a class of laboratory equipment used in the field of biochemistry and molecular biology. These inhibitors are designed to specifically target and inactivate proteases, which are enzymes that break down proteins. Protease inhibitors play a crucial role in various experimental and analytical procedures, such as protein extraction, purification, and stabilization.

Automatically generated - may contain errors

Lab products found in correlation

Phosphatase inhibitor, by Roche (248 mentions) Pvdf membrane, by Merck Group (214 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (210 mentions) Bca protein assay kit, by Thermo Fisher Scientific (198 mentions) Ripa buffer, by Thermo Fisher Scientific (168 mentions) Ripa buffer, by Merck Group (133 mentions) Phosphatase inhibitor, by Merck Group (113 mentions) Ripa lysis buffer, by Beyotime (107 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (78 mentions) Pvdf membrane, by Bio-Rad (76 mentions) Bca assay, by Thermo Fisher Scientific (74 mentions) Nitrocellulose membrane, by Bio-Rad (73 mentions) β actin, by Merck Group (71 mentions) Bradford assay, by Bio-Rad (67 mentions) Anti β actin, by Merck Group (65 mentions) Image lab software, by Bio-Rad (64 mentions) Ripa buffer, by Beyotime (51 mentions) β actin, by Cell Signaling Technology (50 mentions) Polyvinylidene difluoride membrane, by Merck Group (50 mentions) Polyvinylidene fluoride membrane, by Merck Group (50 mentions)

4 287 protocols using protease inhibitor

Tissue Digestion and EC Enrichment Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lymph node and brain tissues were digested with 2.5 mg/mL Collagenase D (Roche), 50 μg/mL DNAse I (Roche), and 0.4× protease inhibitor (Roche) in digestion buffer at 37 °C. Digestion of LNs was adapted from Fletcher et al. [29 (link)], such that tissues were digested for 20 min without protease inhibitor followed by three 10-min incubations with protease inhibitor. Brain tissues were digested for 15–20 min at 37 °C. Enrichment of ECs was performed by centrifugation at 5000 g for 30 min at 4 °C in a dextran solution (17% dextran (Sigma, catalog number 31392)/20 mM HEPES). Skin, colon, and small intestine tissues were digested with 2.5 mg/mL Collagenase D (Roche), 50 μg/mL DNAse I (Roche), and 1× protease inhibitor (Roche) in digestion buffer for 30 min at 37 °C on a rotisserie wheel. Colon and small intestine were washed with 5% FBS and 25 mM HEPES, followed by 2 mM EDTA and 25 mM HEPES, and finally 10% FBS, 5 mM EDTA, and 15 mM HEPES prior to enzymatic digestion to remove epithelial cells. Pancreatic and adipose tissues were digested with 1.25 mg/mL Collagenase D (Roche), 50 μg/mL DNAse I (Roche), and 1× protease inhibitor (Roche) in digestion buffer for 30 min at 37 °C on a rotisserie wheel. Cells were resuspended in FACS buffer (PBS (Lonza), 5% FBS (Invitrogen-Gibco), 5 mM EDTA (Boston BioProducts)) for analysis.

Thiriot A., Perdomo C., Cheng G., Novitzky-Basso I., McArdle S., Kishimoto J.K., Barreiro O., Mazo I., Triboulet R., Ley K., Rot A, & von Andrian U.H. (2017). Differential DARC/ACKR1 expression distinguishes venular from non-venular endothelial cells in murine tissues. BMC Biology, 15, 45.

+ Open protocol

+ Expand

Isolation of Nuclear Proteins from HepG2 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Nuclear protein was isolated from HepG2 cells as previously described (Denison et al., 2002 ) with a few modifications. Briefly, HepG2 cells were treated with vehicle (0.01% DMSO) or TCDD (30 nM in DMSO) for 2h at 37°C Cells were washed twice with 10 mM HEPES (pH 7.5), incubated at 37°C for 15 min, then harvested in MDH buffer [2 mM MgCl₂, 1 mM DTT, 2mM HEPES, pH 7.5, with protease inhibitor (Roche, Indianapolis, IN)] and homogenized using a Dounce homogenizer. The homogenates were centrifuged at 1000g for 5 min, washed twice with MDHK buffer [2 mM MgCl₂, 1 mM DTT, 2mM HEPES, pH 7.5, and 100 mM KCl, with protease inhibitor (Roche)] and centrifuged. The crude nuclear pellets were resuspended in HEDGK4 buffer [25 mM HEPES, pH 7.5, 1 mM EDTA, 1 mM DTT, 10% (v/v) glycerol, and 400 mM KCl, 200 µM phenylmethylsulfonyl fluoride, with protease inhibitor (Roche)] (50 µl per plate of confluent cell) and incubated on ice for 30 min for high-salt extraction followed by centrifugation at 14,000 x g, 4°C for 15 min. The supernatants were ultracentrifuged at 99,000 x g, 4°C for 1 h. The protein concentration in supernatants was determined using the BCA assay (Sigma, St Louis, MO).

Phadnis-Moghe A.S., Li J., Crawford R.B, & Kaminski N.E. (2016). SHP-1 is Directly Activated by the Aryl Hydrocarbon Receptor and regulates BCL-6 in the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Toxicology and applied pharmacology, 310, 41-50.

+ Open protocol

+ Expand

Synaptosomal Membrane Enrichment from Cortical Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

Synaptosome-enriched membranes from 15 DIV cortical neurons were prepared as described previously with some modifications [67 (link)]. Briefly, cultured neurons were stimulated for 15 min with a mixture of Bic/4AP (50 μM/2.5 mM). After a wash in HBSS, neurons were homogenized by passing 15 to 20 times through 0.25G needle in cold buffer containing 0.32 M sucrose, 10 mM HEPES, 15 mM NaF, 15 mM β-glycerophosphate and protease inhibitors (Roche) (pH 7.4). Samples were maintained at 4°C during all steps of the experiment. Homogenates were cleared at 1,000 g for 10 min to remove nuclei and large debris. The resulting supernatants were spun down at 12,000 g for 20 min to obtain a crude membrane fraction and washed twice in HEPES buffer 4 mM containing 1 mM EDTA, 15 mM NaF, 15 mM β-glycerophosphate and protease inhibitors (Roche) (pH 7.4). The resulting pellet was solubilized in 0.5% Triton X-100, 20 mM HEPES, 100 mM NaCl, 15mM NaF, 15 mM β-glycerophosphate (pH 7.2), containing protease inhibitors (Roche) for 20 min at 4°C with mild agitation and analyzed by western blot.

Laporte M.H., Chi K.I., Caudal L.C., Zhao N., Schwarz Y., Rolland M., Martinez-Hernandez J., Martineau M., Chatellard C., Denarier E., Mercier V., Lemaître F., Blot B., Moutaux E., Cazorla M., Perrais D., Lanté F., Bruns D., Fraboulet S., Hemming F.J., Kirchhoff F, & Sadoul R. (2022). Alix is required for activity-dependent bulk endocytosis at brain synapses. PLoS Biology, 20(6), e3001659.

+ Open protocol

+ Expand

Affinity Purification of Chromatin Remodeler

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse ES cells were grown on 15 cm plates until confluency. Cells were harvested and washed with 1 × PBS. Then, cells were resuspended in buffer 1 (10 mM Tris-HCl pH 7.5, 2 mM MgCl₂, 3 mM CaCl₂, Protease inhibitors (Roche)) and incubated for 20 min at 4 °C. After centrifugation, cells were resuspended in buffer 2 (10 mM Tris-HCl pH 7.5, 2 mM MgCl₂, 3 mM CaCl₂, Protease inhibitors (Roche), 0.5% IGEPAL CA-630, 10% glycerol) and incubated for 10 min at 4 °C. After this, cells were again centrifuged and nuclei were resuspended in buffer 3 (50 mM HEPES-KOH pH 7.3, 200 mM KCl, 3.2 mM MgCl₂, 0,25% Triton, 0.25% NP-40, 0.1% Na-deoxycholate, 1 mM DDT, Protease inhibitors (Roche). 4 µl of benzonase was added to the nuclei suspension and was incubated for 1 h at 4 °C. The resulting nuclear lysate was cleared by centrifugation. For the Smarca5 IP, WT and Smarca5-Flag were added to magnetic anti FLAG M2 beads (Merck, Sigma-Aldrich, M8823) and incubated at 4 °C for 2 h. Beads were subsequently washed four times with buffer4 (50 mM HEPES-KOH pH 7.3, 200 mM KCl, 3.2 mM MgCl₂, 0,25% Triton, 0.25% NP-40, 0.1% Na-deoxycholate, 1 mM DDT) and four times with Tris buffer (20 mM Tris-HCl pH 7.5, 137 mM NaCl).

Matzinger M., Schmücker A., Yelagandula R., Stejskal K., Krššáková G., Berger F., Mechtler K, & Mayer R.L. (2024). Micropillar arrays, wide window acquisition and AI-based data analysis improve comprehensiveness in multiple proteomic applications. Nature Communications, 15, 1019.

+ Open protocol

+ Expand

Cellular Fractionation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For whole cell extracts, cells were lysed into RIPA buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 5 mM EDTA, 10 mM K₂HPO₄, 10% (v/v) glycerol, 1% (v/v) Triton X-100, 0.05% SDS, 1 mM DTT, protease inhibitors (Roche) and debris removed prior to loading. For nuclear extracts, cells were resuspended in hypotonic buffer (100 mM HEPES pH 7.9, 10 mM KCl, 1 mM MgCl₂, 1 mM DTT, 1% TritonX-100 and protease inhibitors (Roche)) and sheared following addition of 0.5% NP-40. Nuclei were pelleted by centrifugation at 3 000 rpm for 5 min at 4°C and were resuspended in hypotonic buffer containing 1 U/μl of benzonase (Merck). Extracts were incubated on ice for 30 min, before addition of 400 mM NaCl and incubation for a further 1 h at 4°C, prior to centrifugation at 13 000 rpm for 15 min at 4°C and using the supernatant for analysis. For chromatin extracts, nuclei were harvested as above and the nuclear pellet resuspended in 100 μl extraction buffer (3 mM EDTA, 0.2 mM EGTA and protease inhibitors (Roche)). Samples were spun at 13 000 rpm for 5 min at 4°C and the chromatin pellet resuspended in 20 μl 0.2 M HCl before neutralization with 80 μl 1 M Tris–HCl pH 8.

Kollárovič G., Topping C.E., Shaw E.P, & Chambers A.L. (2019). The human HELLS chromatin remodelling protein promotes end resection to facilitate homologous recombination and contributes to DSB repair within heterochromatin. Nucleic Acids Research, 48(4), 1872-1885.

+ Open protocol

+ Expand

Brain Membrane Protein Extraction Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

All homogenization and centrifugation steps were carried out on ice and at 4°C, respectively. Brain tissues were homogenized in an ice-cold homogenization buffer [10 mM HEPES, pH 7.5, 300 mM sucrose, protease inhibitors (Roche)] using a Dounce homogenizer; the homogenate was centrifuged at 1000 × g for 10 min to remove cell debris and nuclei and the supernatant was collected. The pellet was resuspended again in the homogenization buffer and centrifuged at 1000 × g for 10 min. The pooled supernatants were then centrifuged at 100000 × g for 60 min to enrich membranes. The resulting membrane pellets were washed in 10 mM HEPES, pH 7.5, protease inhibitors (Roche) and solubilized in 50 mM TEAB buffer (Sigma–Aldrich), 7 M urea, 2 M thiourea, 4% CHAPS, 100 mM DTT and protease inhibitors (Roche). The protein concentration was determined by Pierce^TM 660 nm Protein Assay (Thermo Scientific).

Smidak R., Sialana F.J., Kristofova M., Stojanovic T., Rajcic D., Malikovic J., Feyissa D.D., Korz V., Hoeger H., Wackerlig J., Mechtcheriakova D, & Lubec G. (2017). Reduced Levels of the Synaptic Functional Regulator FMRP in Dentate Gyrus of the Aging Sprague-Dawley Rat. Frontiers in Aging Neuroscience, 9, 384.

+ Open protocol

+ Expand

Subcellular Fractionation of Cell Lysates

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The subcellular fractionation protocol was adapted from Batki and colleagues (Batki et al., 2019 (link)). Cell pellets resuspended in LB1 (10 mM Tris-HCl pH 7.5, 2 mM MgCl₂, 3 mM CaCl₂ supplemented with protease inhibitors [Roche]) for 5 min at 4°C followed by centrifugation (1000 g, 5 min, 4°C). The supernatant was saved (swell fraction) and the cell pellet was resuspended and incubated in LB2 (10 mM Tris-HCl pH 7.5, 2 mM MgCl₂, 3 mM CaCl₂0.5% Nonidet P-40% and 10% glycerol, supplemented with protease inhibitors (Roche)) for 5 min at 4°C followed by another centrifugation step (1000 g, 5 min, 4°C). The supernatant was saved (cytoplasmic fraction) and the nuclei pellet was resuspended in LB3 (50 mM Tris-HCl pH 8, 150 mM NaCl, 2 mM MgCl₂, 0.5% Triton X-100, 0.25% Nonidet P-40% and 10% glycerol, supplemented with protease inhibitors [Roche]). After a 10-min incubation on ice, the nuclear lysate was sonicated for 3 cycles of 30 s ON/30 s OFF using a Bioruptor Pico (Diagenode) to ensure solubilisation of chromatin and then spun 18,000 g for 10 min. The swell and cytoplasmic fraction was combined and the fractions were analysed by western blotting.

Eastwood E.L., Jara K.A., Bornelöv S., Munafò M., Frantzis V., Kneuss E., Barbar E.J., Czech B, & Hannon G.J. (2021). Dimerisation of the PICTS complex via LC8/Cut-up drives co-transcriptional transposon silencing in Drosophila. eLife, 10, e65557.

+ Open protocol

+ Expand

Isolation and Analysis of CRY1-YFP and UVR8-YFP Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols

After infiltration for 48 h, tobacco leaves coexpressing Nuc2-COP1-TAP and CRY1-YFP or UVR8-YFP, Nuc2-TAP, and CRY1-YFP or UVR8-YFP were ground in liquid nitrogen, and total proteins were extracted as described (Lee et al., 2007 (link)) using extraction buffer (0.4 M sucrose, 10 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 5 mM β-mercaptoethanol, 0.1 mM phenylmethylsulfonyl fluoride, and 1× protease inhibitor [Roche, Basel, Switzerland]). They were filtered through four-layer Miracloth (Millipore) twice, and 2-ml samples were taken to incubate with 60 μl of anti-GFP magnetic beads (MBL, Japan) at 4°C for 2 h to capture proteins associated with CRY1-YFP or UVR8-YFP. The beads were washed five times with wash buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 10% glycerol, 1 mM EDTA, 0.01% Triton X-100, 0.1 mM phenylmethylsulfonyl fluoride, and 1× protease inhibitor [Roche]). The bound proteins were eluted by heating at 95°C for 10 min and analyzed by Western blot using anti-GFP (Sigma-Aldrich) and anti-TAP (Sigma-Aldrich) antibodies, respectively.

Liu Y., Liu Q., Yan Q., Shi L, & Fang Y. (2014). Nucleolus-tethering system (NoTS) reveals that assembly of photobodies follows a self-organization model. Molecular Biology of the Cell, 25(8), 1366-1373.

+ Open protocol

+ Expand

Co-Immunoprecipitation of RNA Polymerase Subunits

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were harvested in RIPA lysis buffer supplemented with protease inhibitors (Roche). Protein lysates (1 mg) were precleared using Dynabeads Protein G beads (Invitrogen 10003D) for 1 hour at 4˚C and centrifuged at 5,000 rpm for 5 minutes at 4°C. Primary antibodies (2 μg) against RPA194 (C-1; Santa Cruz Biotechnology), RPA135 (H-15; Santa Cruz Biotechnology) or control anti-mouse IgG (Millipore Sigma) were incubated with the supernatant overnight with rotation at 4˚C. The protein-antibody mixture was collected on Dynabeads Protein G beads (50 μL per sample) for 45 minutes at 4°C followed by 5 washes with RIPA buffer with protease inhibitor (Roche). The beads were resuspended in 2x Laemmli Sample Buffer with DTT and boiled for 10 minutes. Samples were run on SDS-PAGE gel and transferred to PVDF membranes (Millipore Sigma) followed by immunoblotting.

Ford B.L., Wei T., Liu H., Scull C.E., Najmi S.M., Pitts S., Fan W., Schneider D.A, & Laiho M. (2023). Expression of RNA polymerase I catalytic core is influenced by RPA12. PLOS ONE, 18(5), e0285660.

+ Open protocol

+ Expand

Brassinin's Impact on Cancer Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The PC-3, DU145, and LNCaP cells were exposed to various concentrations of Brassinin for 48 h, lysed with lysis solution (150 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM EDTA 50 mM Tris–HCl, pH 7.4, 1 mM Na3VO4, 1 mM NaF, and 1 × protease inhibitor cocktail) containing protease inhibitors (Roche Diagnostics GmbH, Mannheim, Germany) and phosphatase inhibitors (SigmaAldrich, St. Louis, MO, USA) on ice, and centrifuged at 13,000 rpm for 20 min at 4 °C. The cell lysates were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a Hybond ECL transfer membrane. The membranes were blocked using 5% skim milk and incubated with primary antibodies for Caspase3, PARP, Bcl-2, Glut1, HK2, PKM2, LDH, SIRT1, β-catenin, c-Myc (Cell signaling Technology, Beverly, MA, USA), and β-actin (Sigma, St. Louis, MO, USA). The washed membranes were incubated with the corresponding horseradish peroxidase-conjugated anti-rabbit IgG (dilution 1:5000, cat. no. 7074; Cell Signaling Technology, Inc.) and anti-mouse IgG (dilution 1:5000, cat. no. 7076; Cell Signaling Technology, Danvers, MA, USA). The immunoreactive polypeptides were analyzed using an enhanced chemiluminescence (Thermo Scientific, Rochester, NY, USA).

Kwon H.H., Ahn C.H., Lee H.J., Sim D.Y., Park J.E., Park S.Y., Kim B., Shim B.S, & Kim S.H. (2023). The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis. International Journal of Molecular Sciences, 24(18), 13912.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!