protease cocktail inhibitors  (Millipore)


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    Structured Review

    Millipore protease cocktail inhibitors
    Protease Cocktail Inhibitors, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease cocktail inhibitors/product/Millipore
    Average 90 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    protease cocktail inhibitors - by Bioz Stars, 2020-03
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: The gene was cloned into the pET21a vector, and various E. coli strains and growing temperatures were screened for optimum soluble protein production. .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water).

    Centrifugation:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: After an incubation period of 72 h, cells were collected by centrifugation at 2500 rpm for 7 min, washed twice with PBS and finally resuspended in 300 μl of PBS containing 2.5 μM of annexin V-FITC (Thermo Fisher, MA, USA) and 1 μg ml-1 propidium iodide (Sigma-Aldrich) for 10 min at 37°C in the dark. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water). .. After centrifugation at 30,000 × g for 30 min at 4 °C, the supernatant was loaded onto an Ni-affinity column previously equilibrated with 20 mM sodium phosphate at pH 8, 150 mM NaCl, 0.1 mM DTT, and 10 mM imidazole.

    Synthesized:

    Article Title: Cetuximab-conjugated iron oxide nanoparticles for cancer imaging and therapy
    Article Snippet: The cells were starved in starvation medium (culture medium containing 0.5% fetal bovine serum) for 24 hours at 37°C and then treated under serum starvation conditions for 2 hours with cetuximab and the synthesized SPIONs prior to stimulation with 30 ng/mL epidermal growth factor for 30 minutes at 37°C. .. After washing once with cold 1× PBS, the cell pellet was lysed with RIPA lysis buffer (1× PBS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM sodium fluoride, 0.25% sodium deoxycholate, 1% NP-40, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride) with 1% freshly added SigmaFAST™ protease inhibitor cocktail (s8830, Sigma-Aldrich, St Louis, MO, USA).

    Cell Fractionation:

    Article Title: Insect Stage-Specific Receptor Adenylate Cyclases Are Localized to Distinct Subdomains of the Trypanosoma brucei Flagellar Membrane
    Article Snippet: Paragraph title: Cell fractionation and immunoblotting. ... To assay for an association with detergent-resistant membranes (see ), cells were washed once in PBS and lysed at either 4°C or 37°C in PBS with 1% Triton X-100 and protease inhibitors (SigmaFAST cocktail; Sigma-Aldrich) as described previously ( ).

    Cytometry:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: Apoptotic cells were determined by analyzing 60,000 ungated cells by using a flow cytometer (Coulter Cytoflex, Beckman, CA, USA) and the FlowJo software. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Blocking Assay:

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. .. Membranes were blocked in Licor Odyssey blocking buffer, incubated with primary antibody diluted (1:1000 for anti-V5 or 1:2000 for anti-PfLDH) in blocking buffer, and then incubated with secondary antibodies diluted in blocking buffer.

    Incubation:

    Article Title: MBNL splicing activity depends on RNA binding site structural context
    Article Snippet: Immunoblotting HeLa cells were lysed with RIPA buffer supplemented with SIGMAFAST Protease Inhibitor Cocktail (Sigma). .. Membranes were blocked for 1 h with 5% skim milk in PBST buffer (phosphate-buffered saline (PBS), 0.1% Tween-20) and incubated with a primary antibody against eGFP (1:1000, Santa Cruz cat. no. sc-8334) or human GAPDH (1:1000, Santa Cruz cat. no. sc-47724).

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. .. Membranes were blocked in Licor Odyssey blocking buffer, incubated with primary antibody diluted (1:1000 for anti-V5 or 1:2000 for anti-PfLDH) in blocking buffer, and then incubated with secondary antibodies diluted in blocking buffer.

    Article Title: Cetuximab-conjugated iron oxide nanoparticles for cancer imaging and therapy
    Article Snippet: After washing once with cold 1× PBS, the cell pellet was lysed with RIPA lysis buffer (1× PBS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM sodium fluoride, 0.25% sodium deoxycholate, 1% NP-40, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride) with 1% freshly added SigmaFAST™ protease inhibitor cocktail (s8830, Sigma-Aldrich, St Louis, MO, USA). .. The membranes were then incubated with primary antibodies against phospho-EGFR (Tyr1173; 1:2,000; Novus Biologi-cals Inc, Littleton, CO, USA) and β-actin (1:10,000; GeneTex Inc, Irvine, CA, USA), followed by horseradish peroxidase-conjugated goat anti-rabbit IgG (H + L) secondary antibodies (1:10,000; Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA).

    Article Title: Stimulation of Dectin-1 and Dectin-2 during Parenteral Immunization, but Not Mincle, Induces Secretory IgA in Intestinal Mucosa
    Article Snippet: For collection of intestinal lavage (IL) samples, small intestine was removed and carefully filled with 3 mL of PBS containing Sigmafast Protease Inhibitor Cocktail (Sigma, USA) using a syringe. .. After 10 min incubation, solution was transferred into the 1.5 mL conical tubes (Eppendorf, Germany).

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich). ..

    Expressing:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: To investigate the effect of nutlin-3a and of Nut-Mag-SLNs on the expression of p53 and its downstream proteins (e.g., MDM2 and p21), western blotting analysis was performed. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: The best expression conditions were E. coli Rosetta (DE3) cells and overnight induction with 0.5 mM isopropyl-β- d -1-thiogalactopyranoside (IPTG) at 24 °C. .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water).

    BIA-KA:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich). .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Modification:

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: .. Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO). .. Samples were warmed to 55°C, and sonicated 4 × 30 seconds in a Bioruptor (Diagenode, Denville, NJ).

    Article Title: Surfen, a proteoglycan binding agent, reduces inflammation but inhibits remyelination in murine models of Multiple Sclerosis
    Article Snippet: .. Acetic acid, ammonium sulphamate (H6 N2 O3 S), bovine serum albumin (BSA), brefeldin A from penicillium , complete Freund’s adjuvant (CFA) containing mycobacterium tuberculosis H37RA (10 mg/ml), concanavalin A, dimethyl sulfoxide (DMSO), chondroitinase ABC, cysteine hydrochloric acid (HCL), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), eriochrome cyanine, Dulbecco’s Modified Eagle’s Medium (DMEM), 10% buffered formalin, Griess reagent for nitrite, ketoprofen, L-α-Lysophosphatidylcholine (lysolecithin) from egg yolk, phosphate buffered saline (PBS; pH 7.4), pertussis toxin from Bordetella pertussis, Roswell Park Memorial Institute 1640 medium (RPMI), heparinase-III, ionomycin, bacterial lipopolysaccharide (LPS) from E. coli 0111:B4, papain from papaya latex, phorbol 12-myristate 13-acetate (PMA), sodium azide (NaN3 ), sodium citrate, sodium nitrite (NaNO2 ), sodium phosphate (NaH2 PO4 ), SIGMA FAST ® protease inhibitor cocktail tablets, surfen (bis-2-methyl-4-amino-quinolyl-6-carbamide hydrate), and Triton-X-100 were obtained from Sigma-Aldrich Canada (Oakville, ON). .. Fetal bovine serum (FBS), 10,000 U/ml penicillin/10,000 μg/mL streptomycin solution, 200 mM L-glutamine, 1 M 4-(2-hydroyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution, and 0.4% trypan blue dye solution were obtained from Invitrogen Canada (Oakville, ON).

    Western Blot:

    Article Title: MBNL splicing activity depends on RNA binding site structural context
    Article Snippet: Immunoblotting HeLa cells were lysed with RIPA buffer supplemented with SIGMAFAST Protease Inhibitor Cocktail (Sigma). .. Anti-rabbit (1:20 000, Sigma cat. no. A9169) and anti-mouse (1:2000, Millipore cat. no. 12-349) secondary antibodies were conjugated with horseradish peroxidase and detected using the Pierce ECL Plus Western Blotting Substrate detection kit (Thermo Scientific).

    Article Title: Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p
    Article Snippet: Paragraph title: Western blot ... The lysis buffer was Radio Immunoprecipitation Assay (RIPA) buffer with SIGMAFAST Protease Inhibitor Cocktail Tablets (Sigma-Aldrich, St. Louis, USA).

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: .. Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. ..

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: Paragraph title: MCL1 and MRPS12 protein extraction and western blotting ... Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO).

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: To investigate the effect of nutlin-3a and of Nut-Mag-SLNs on the expression of p53 and its downstream proteins (e.g., MDM2 and p21), western blotting analysis was performed. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Article Title: The Distribution of Cholinergic Neurons, and their Co-localization with FMRFamide in Central and Peripheral Neurons of the Spider Cupiennius salei
    Article Snippet: Paragraph title: Western blot analysis ... Spider brain tissue was placed in a 1 ml glass tissue grinder containing ice-cold spider saline with protease inhibitor (Sigma S-8830) at a dilution of 1 part protease inhibitor to 10 parts spider saline.

    Article Title: Extracellular Vesicles Released by Oxidatively Injured or Intact C2C12 Myotubes Promote Distinct Responses Converging toward Myogenesis
    Article Snippet: Paragraph title: 4.6. Western Blotting Analysis ... The protein pellet was dissolved in ISOT buffer (8 M urea, 4% CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate), 65 mM DTE (1,4-Dithioerythritol), 40 mM Tris base and added with SIGMAFAST™ Protease Inhibitor Cocktail (Sigma-Aldrich, Milan, Italy)) and sonicated for 5 s on ice.

    Flow Cytometry:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: Apoptotic cells were determined by analyzing 60,000 ungated cells by using a flow cytometer (Coulter Cytoflex, Beckman, CA, USA) and the FlowJo software. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Serial Dilution:

    Article Title: Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p
    Article Snippet: The cells were treated with a serial dilution of miR-181d-5p mimics or inhibitors. .. The lysis buffer was Radio Immunoprecipitation Assay (RIPA) buffer with SIGMAFAST Protease Inhibitor Cocktail Tablets (Sigma-Aldrich, St. Louis, USA).

    Radio Ligand Binding Assay:

    Article Title: Structural basis of ligand binding modes at the neuropeptide Y Y1 receptor
    Article Snippet: Total solublized protein of Sf 9 membrane preparations (see above) used in radio ligand binding assay was determined by the Bradford method according to the manufacturers’ protocol (BioRad Protein Assay; BioRad, Munich, Germany). .. Aliquots of homogenized membrane preparations, corresponding to 100 μg of protein, were centrifuged at 50,000 g at 4 °C for 15 min, and the pellets were re-suspended in 50 mM Tris, pH 7.4, supplemented with 1 mM EDTA and protease inhibitors (SIGMAFAST Protease Inhibitor cocktail tablets, Sigma) at a protein concentration of 1,600 μg ml−1 .

    Protein Concentration:

    Article Title: Structural basis of ligand binding modes at the neuropeptide Y Y1 receptor
    Article Snippet: .. Aliquots of homogenized membrane preparations, corresponding to 100 μg of protein, were centrifuged at 50,000 g at 4 °C for 15 min, and the pellets were re-suspended in 50 mM Tris, pH 7.4, supplemented with 1 mM EDTA and protease inhibitors (SIGMAFAST Protease Inhibitor cocktail tablets, Sigma) at a protein concentration of 1,600 μg ml−1 . .. Membrane homogenates (15 μl) were processed and subjected to immunoblotting as described previously with the following modifications: blotting onto the nitrocellulose membrane was performed at 60 mA for 60 min. Primary antibody ANTI-FLAG M1 from mouse (Sigma, order no. F3040, lot SLBK1592V) was diluted 1:500.

    Article Title: Cetuximab-conjugated iron oxide nanoparticles for cancer imaging and therapy
    Article Snippet: After washing once with cold 1× PBS, the cell pellet was lysed with RIPA lysis buffer (1× PBS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM sodium fluoride, 0.25% sodium deoxycholate, 1% NP-40, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride) with 1% freshly added SigmaFAST™ protease inhibitor cocktail (s8830, Sigma-Aldrich, St Louis, MO, USA). .. The resulting mixture was centrifuged at 14,000× g for 10 minutes at 4°C, and the protein concentration in the supernatant was measured using Bradford reagents (Sigma-Aldrich).

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO). .. Protein concentration was determined using bicinchoninic acid microplate assay (Thermo Scientific, Rockford, IL).

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich). .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Sonication:

    Article Title: Disease‐associated tau impairs mitophagy by inhibiting Parkin translocation to mitochondria
    Article Snippet: Animals were resuspended in RIPA buffer [10 mM Tris–HCl (pH 7.4), 140 mM NaCl, 1% Triton X‐100, 0.1% sodium deoxycholate and 0.1% SDS, 1 mM EDTA] supplemented with protease inhibitors (Sigma, S8830). .. The animals were lysed by douncing followed by sonication for 30 s, five times (amplitude: 40%; Sonics VibraCell VCX130).

    Article Title: Stimulation of Dectin-1 and Dectin-2 during Parenteral Immunization, but Not Mincle, Induces Secretory IgA in Intestinal Mucosa
    Article Snippet: For collection of intestinal lavage (IL) samples, small intestine was removed and carefully filled with 3 mL of PBS containing Sigmafast Protease Inhibitor Cocktail (Sigma, USA) using a syringe. .. Tubes were placed in ice and sonicated for 20 minutes in ultrasonic water bath (Elma Schmidbauer, Germany).

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO). .. Samples were warmed to 55°C, and sonicated 4 × 30 seconds in a Bioruptor (Diagenode, Denville, NJ).

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water). .. After centrifugation at 30,000 × g for 30 min at 4 °C, the supernatant was loaded onto an Ni-affinity column previously equilibrated with 20 mM sodium phosphate at pH 8, 150 mM NaCl, 0.1 mM DTT, and 10 mM imidazole.

    Article Title: Extracellular Vesicles Released by Oxidatively Injured or Intact C2C12 Myotubes Promote Distinct Responses Converging toward Myogenesis
    Article Snippet: .. The protein pellet was dissolved in ISOT buffer (8 M urea, 4% CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate), 65 mM DTE (1,4-Dithioerythritol), 40 mM Tris base and added with SIGMAFAST™ Protease Inhibitor Cocktail (Sigma-Aldrich, Milan, Italy)) and sonicated for 5 s on ice. ..

    MTT Assay:

    Article Title: Surfen, a proteoglycan binding agent, reduces inflammation but inhibits remyelination in murine models of Multiple Sclerosis
    Article Snippet: .. Acetic acid, ammonium sulphamate (H6 N2 O3 S), bovine serum albumin (BSA), brefeldin A from penicillium , complete Freund’s adjuvant (CFA) containing mycobacterium tuberculosis H37RA (10 mg/ml), concanavalin A, dimethyl sulfoxide (DMSO), chondroitinase ABC, cysteine hydrochloric acid (HCL), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), eriochrome cyanine, Dulbecco’s Modified Eagle’s Medium (DMEM), 10% buffered formalin, Griess reagent for nitrite, ketoprofen, L-α-Lysophosphatidylcholine (lysolecithin) from egg yolk, phosphate buffered saline (PBS; pH 7.4), pertussis toxin from Bordetella pertussis, Roswell Park Memorial Institute 1640 medium (RPMI), heparinase-III, ionomycin, bacterial lipopolysaccharide (LPS) from E. coli 0111:B4, papain from papaya latex, phorbol 12-myristate 13-acetate (PMA), sodium azide (NaN3 ), sodium citrate, sodium nitrite (NaNO2 ), sodium phosphate (NaH2 PO4 ), SIGMA FAST ® protease inhibitor cocktail tablets, surfen (bis-2-methyl-4-amino-quinolyl-6-carbamide hydrate), and Triton-X-100 were obtained from Sigma-Aldrich Canada (Oakville, ON). .. Fetal bovine serum (FBS), 10,000 U/ml penicillin/10,000 μg/mL streptomycin solution, 200 mM L-glutamine, 1 M 4-(2-hydroyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution, and 0.4% trypan blue dye solution were obtained from Invitrogen Canada (Oakville, ON).

    Fluorescence:

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. .. Membranes were scanned on a Licor Odyssey CLx imager system and quantified using volumetric measurement of fluorescence intensity.

    Isolation:

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water). .. After centrifugation at 30,000 × g for 30 min at 4 °C, the supernatant was loaded onto an Ni-affinity column previously equilibrated with 20 mM sodium phosphate at pH 8, 150 mM NaCl, 0.1 mM DTT, and 10 mM imidazole.

    Mouse Assay:

    Article Title: Stimulation of Dectin-1 and Dectin-2 during Parenteral Immunization, but Not Mincle, Induces Secretory IgA in Intestinal Mucosa
    Article Snippet: Two weeks after the second immunization, mice were euthanized by CO2 overdose. .. For collection of intestinal lavage (IL) samples, small intestine was removed and carefully filled with 3 mL of PBS containing Sigmafast Protease Inhibitor Cocktail (Sigma, USA) using a syringe.

    Protein Extraction:

    Article Title: Disease‐associated tau impairs mitophagy by inhibiting Parkin translocation to mitochondria
    Article Snippet: Paragraph title: C. elegans protein extraction and quantification ... Animals were resuspended in RIPA buffer [10 mM Tris–HCl (pH 7.4), 140 mM NaCl, 1% Triton X‐100, 0.1% sodium deoxycholate and 0.1% SDS, 1 mM EDTA] supplemented with protease inhibitors (Sigma, S8830).

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: Paragraph title: MCL1 and MRPS12 protein extraction and western blotting ... Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO).

    Purification:

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: .. Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. ..

    SDS Page:

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO). .. Samples were mixed 1:4 with 5x Loading Buffer Supplement (50% glycerol, 0.02% bromphenol blue, 12.5% β-mercaptoethanol), separated by SDS-PAGE on NuPage Novex 4-12% Bis-Tris Midi gels (Life Technologies, Grand Island, NY), and transferred to Immobilon Fl (Millipore, Billerica, MA) PVDF membranes.

    Plasmid Preparation:

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: The gene was cloned into the pET21a vector, and various E. coli strains and growing temperatures were screened for optimum soluble protein production. .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water).

    Software:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: Apoptotic cells were determined by analyzing 60,000 ungated cells by using a flow cytometer (Coulter Cytoflex, Beckman, CA, USA) and the FlowJo software. .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    In Vitro:

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: Paragraph title: Cell cultures &in vitro testing ... After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich).

    Radio Immunoprecipitation:

    Article Title: Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p
    Article Snippet: .. The lysis buffer was Radio Immunoprecipitation Assay (RIPA) buffer with SIGMAFAST Protease Inhibitor Cocktail Tablets (Sigma-Aldrich, St. Louis, USA). ..

    Protease Inhibitor:

    Article Title: MBNL splicing activity depends on RNA binding site structural context
    Article Snippet: .. Immunoblotting HeLa cells were lysed with RIPA buffer supplemented with SIGMAFAST Protease Inhibitor Cocktail (Sigma). .. Samples were heated to 95°C for 5 min, separated on 10% SDS polyacrylamide gels and transferred to nitrocellulose (Protran BA 85, Whatman) using a wet transfer apparatus (1 h, 100 V, 4°C).

    Article Title: Aberrant activity of NKL homeobox gene NKX3-2 in a T-ALL subset
    Article Snippet: .. Protein lysates from cell lines were prepared using SIGMAFast protease inhibitor cocktail (Sigma). .. Cell lysates from primary human spleen were obtained from Origene.

    Article Title: Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p
    Article Snippet: .. The lysis buffer was Radio Immunoprecipitation Assay (RIPA) buffer with SIGMAFAST Protease Inhibitor Cocktail Tablets (Sigma-Aldrich, St. Louis, USA). ..

    Article Title: Structural basis of ligand binding modes at the neuropeptide Y Y1 receptor
    Article Snippet: .. Aliquots of homogenized membrane preparations, corresponding to 100 μg of protein, were centrifuged at 50,000 g at 4 °C for 15 min, and the pellets were re-suspended in 50 mM Tris, pH 7.4, supplemented with 1 mM EDTA and protease inhibitors (SIGMAFAST Protease Inhibitor cocktail tablets, Sigma) at a protein concentration of 1,600 μg ml−1 . .. Membrane homogenates (15 μl) were processed and subjected to immunoblotting as described previously with the following modifications: blotting onto the nitrocellulose membrane was performed at 60 mA for 60 min. Primary antibody ANTI-FLAG M1 from mouse (Sigma, order no. F3040, lot SLBK1592V) was diluted 1:500.

    Article Title: An essential contractile ring protein controls cell division in Plasmodium falciparum
    Article Snippet: .. Quantitative western blotting Parasite pellets were purified by lysing RBCs in 0.05% saponin in PBS with protease inhibitors (SigmaFast Protease Inhibitor Cocktail, Sigma) and boiled in Laemmli buffer. ..

    Article Title: Cetuximab-conjugated iron oxide nanoparticles for cancer imaging and therapy
    Article Snippet: .. After washing once with cold 1× PBS, the cell pellet was lysed with RIPA lysis buffer (1× PBS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM sodium fluoride, 0.25% sodium deoxycholate, 1% NP-40, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride) with 1% freshly added SigmaFAST™ protease inhibitor cocktail (s8830, Sigma-Aldrich, St Louis, MO, USA). .. The resulting mixture was centrifuged at 14,000× g for 10 minutes at 4°C, and the protein concentration in the supernatant was measured using Bradford reagents (Sigma-Aldrich).

    Article Title: Stimulation of Dectin-1 and Dectin-2 during Parenteral Immunization, but Not Mincle, Induces Secretory IgA in Intestinal Mucosa
    Article Snippet: .. For collection of intestinal lavage (IL) samples, small intestine was removed and carefully filled with 3 mL of PBS containing Sigmafast Protease Inhibitor Cocktail (Sigma, USA) using a syringe. .. After 10 min incubation, solution was transferred into the 1.5 mL conical tubes (Eppendorf, Germany).

    Article Title: Transcriptomic profiles of aging in purified human immune cells
    Article Snippet: .. Pellet was resuspended in 100 μl modified 4× Laemmli buffer [ ] (4% SDS, 250 mM Tris HCl, no glycerol, no bromophenol blue, no β-mercaptoethanol) mixed 1:1 with 8 M Urea [ ], with SigmaFAST Protease Inhibitor Cocktail Tablet (Sigma-Aldrich, St. Louis, MO). .. Samples were warmed to 55°C, and sonicated 4 × 30 seconds in a Bioruptor (Diagenode, Denville, NJ).

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich). ..

    Article Title: Functional Characterization and Structure-Guided Mutational Analysis of the Transsulfuration Enzyme Cystathionine γ-Lyase from Toxoplasma gondii
    Article Snippet: .. Cells were isolated by centrifugation (5000 g for 15 min), resuspended in 20 mM sodium phosphate pH 8, 150 mM NaCl, and 0.1 mM DTT buffer containing 1× protease inhibitor EDTA free (Sigma-Aldrich, S8830, Milano, Italy), and lysed by sonication (12 cycles of 10 s ON/20 s OFF on ice water). .. After centrifugation at 30,000 × g for 30 min at 4 °C, the supernatant was loaded onto an Ni-affinity column previously equilibrated with 20 mM sodium phosphate at pH 8, 150 mM NaCl, 0.1 mM DTT, and 10 mM imidazole.

    Article Title: Surfen, a proteoglycan binding agent, reduces inflammation but inhibits remyelination in murine models of Multiple Sclerosis
    Article Snippet: .. Acetic acid, ammonium sulphamate (H6 N2 O3 S), bovine serum albumin (BSA), brefeldin A from penicillium , complete Freund’s adjuvant (CFA) containing mycobacterium tuberculosis H37RA (10 mg/ml), concanavalin A, dimethyl sulfoxide (DMSO), chondroitinase ABC, cysteine hydrochloric acid (HCL), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), eriochrome cyanine, Dulbecco’s Modified Eagle’s Medium (DMEM), 10% buffered formalin, Griess reagent for nitrite, ketoprofen, L-α-Lysophosphatidylcholine (lysolecithin) from egg yolk, phosphate buffered saline (PBS; pH 7.4), pertussis toxin from Bordetella pertussis, Roswell Park Memorial Institute 1640 medium (RPMI), heparinase-III, ionomycin, bacterial lipopolysaccharide (LPS) from E. coli 0111:B4, papain from papaya latex, phorbol 12-myristate 13-acetate (PMA), sodium azide (NaN3 ), sodium citrate, sodium nitrite (NaNO2 ), sodium phosphate (NaH2 PO4 ), SIGMA FAST ® protease inhibitor cocktail tablets, surfen (bis-2-methyl-4-amino-quinolyl-6-carbamide hydrate), and Triton-X-100 were obtained from Sigma-Aldrich Canada (Oakville, ON). .. Fetal bovine serum (FBS), 10,000 U/ml penicillin/10,000 μg/mL streptomycin solution, 200 mM L-glutamine, 1 M 4-(2-hydroyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution, and 0.4% trypan blue dye solution were obtained from Invitrogen Canada (Oakville, ON).

    Article Title: Extracellular Vesicles Released by Oxidatively Injured or Intact C2C12 Myotubes Promote Distinct Responses Converging toward Myogenesis
    Article Snippet: .. The protein pellet was dissolved in ISOT buffer (8 M urea, 4% CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate), 65 mM DTE (1,4-Dithioerythritol), 40 mM Tris base and added with SIGMAFAST™ Protease Inhibitor Cocktail (Sigma-Aldrich, Milan, Italy)) and sonicated for 5 s on ice. ..

    Lysis:

    Article Title: Strontium inhibits osteoclastogenesis by enhancing LRP6 and β-catenin-mediated OPG targeted by miR-181d-5p
    Article Snippet: .. The lysis buffer was Radio Immunoprecipitation Assay (RIPA) buffer with SIGMAFAST Protease Inhibitor Cocktail Tablets (Sigma-Aldrich, St. Louis, USA). ..

    Article Title: Cetuximab-conjugated iron oxide nanoparticles for cancer imaging and therapy
    Article Snippet: .. After washing once with cold 1× PBS, the cell pellet was lysed with RIPA lysis buffer (1× PBS, 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM sodium fluoride, 0.25% sodium deoxycholate, 1% NP-40, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride) with 1% freshly added SigmaFAST™ protease inhibitor cocktail (s8830, Sigma-Aldrich, St Louis, MO, USA). .. The resulting mixture was centrifuged at 14,000× g for 10 minutes at 4°C, and the protein concentration in the supernatant was measured using Bradford reagents (Sigma-Aldrich).

    Article Title: Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment
    Article Snippet: .. After 72 h of treatment, cells were centrifuged at 2500 rpm for 7 min, washed with PBS and finally obtained pellets were incubated with RIPA lysis buffer (Sigma-Aldrich) containing serine/threonine and tyrosine phosphatase inhibitors and a protease inhibitor cocktail 1× (S8830, Sigma-Aldrich). ..

    Article Title: Extracellular Vesicles Released by Oxidatively Injured or Intact C2C12 Myotubes Promote Distinct Responses Converging toward Myogenesis
    Article Snippet: Western Blotting Analysis Protein extracts were obtained from the organic phase after Qiazol cell lysis following Qiagen User Protocol RY16 May-04. .. The protein pellet was dissolved in ISOT buffer (8 M urea, 4% CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate), 65 mM DTE (1,4-Dithioerythritol), 40 mM Tris base and added with SIGMAFAST™ Protease Inhibitor Cocktail (Sigma-Aldrich, Milan, Italy)) and sonicated for 5 s on ice.

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  • 99
    Millipore protease inhibitor cocktail iii
    The protein complementation assay confirms the effects of the R282H mutation and a phosphorylation site mutation on TIN2L interaction with TRF2. (A) Quantification of fluorescence from coexpression of V1-TRF2 with TIN2L-V2, TIN2L-D391K+D395K-V2, or TIN2L-R282H-V2. Error bars represent the <t>SDs</t> from <t>three</t> separate transfections each measured in triplicate. *, P
    Protease Inhibitor Cocktail Iii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease inhibitor cocktail iii/product/Millipore
    Average 99 stars, based on 79 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail iii - by Bioz Stars, 2020-03
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    99
    Millipore protease inhibitor cocktail set iii
    Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) <t>BAL,</t> ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of <t>three</t> independent experiments. * P
    Protease Inhibitor Cocktail Set Iii, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 107 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protease inhibitor cocktail set iii/product/Millipore
    Average 99 stars, based on 107 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail set iii - by Bioz Stars, 2020-03
    99/100 stars
      Buy from Supplier

    Image Search Results


    The protein complementation assay confirms the effects of the R282H mutation and a phosphorylation site mutation on TIN2L interaction with TRF2. (A) Quantification of fluorescence from coexpression of V1-TRF2 with TIN2L-V2, TIN2L-D391K+D395K-V2, or TIN2L-R282H-V2. Error bars represent the SDs from three separate transfections each measured in triplicate. *, P

    Journal: Molecular and Cellular Biology

    Article Title: The C-Terminal Extension Unique to the Long Isoform of the Shelterin Component TIN2 Enhances Its Interaction with TRF2 in a Phosphorylation- and Dyskeratosis Congenita Cluster-Dependent Fashion

    doi: 10.1128/MCB.00025-18

    Figure Lengend Snippet: The protein complementation assay confirms the effects of the R282H mutation and a phosphorylation site mutation on TIN2L interaction with TRF2. (A) Quantification of fluorescence from coexpression of V1-TRF2 with TIN2L-V2, TIN2L-D391K+D395K-V2, or TIN2L-R282H-V2. Error bars represent the SDs from three separate transfections each measured in triplicate. *, P

    Article Snippet: Cells were resuspended in ice-cold lysis buffer (50 mM Tris-HCl at pH 7.5, 1 mM EDTA, 400 mM NaCl, 1% Triton X-100, 0.1% SDS, 1 mM dithiothreitol [DTT], 1 mM phenylmethylsulfonyl fluoride [PMSF], and 1× protease inhibitor cocktail III [Calbiochem]) and incubated for 10 min on ice prior to addition of an equal amount of ice-cold water.

    Techniques: Mutagenesis, Fluorescence, Transfection

    Etf-1 interacts with the RAB5-class III PtdIns3K complex. Co-immunoprecipitation of native Etf-1 with endogenous RAB5, BECN1, and PIK3C3/VPS34. Uninfected (THP) or E. chaffeensis -infected ( Ech ) THP-1 cells at 2 d p.i. were lysed in modified lysis buffer and immunoprecipitated ( IP ) with rabbit anti-Etf-1, BECN1, or PIK3C3, or mouse anti-RAB5 IgG cross-linked to protein A/G-magnetic beads for 2 h. Bound proteins were eluted and subjected to western blotting. Arrows indicate the target proteins. *, IgG heavy or light chains. The number under the figure is the density ratio relative to ACT/actin, with uninfected cells set as 1. The absence of a number indicates an infinite ratio (i.e., the protein was absent in the control). Images were representative of 3 experiments with similar results.

    Journal: Autophagy

    Article Title: Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase

    doi: 10.1080/15548627.2016.1217369

    Figure Lengend Snippet: Etf-1 interacts with the RAB5-class III PtdIns3K complex. Co-immunoprecipitation of native Etf-1 with endogenous RAB5, BECN1, and PIK3C3/VPS34. Uninfected (THP) or E. chaffeensis -infected ( Ech ) THP-1 cells at 2 d p.i. were lysed in modified lysis buffer and immunoprecipitated ( IP ) with rabbit anti-Etf-1, BECN1, or PIK3C3, or mouse anti-RAB5 IgG cross-linked to protein A/G-magnetic beads for 2 h. Bound proteins were eluted and subjected to western blotting. Arrows indicate the target proteins. *, IgG heavy or light chains. The number under the figure is the density ratio relative to ACT/actin, with uninfected cells set as 1. The absence of a number indicates an infinite ratio (i.e., the protein was absent in the control). Images were representative of 3 experiments with similar results.

    Article Snippet: Co-immunoprecipitation Uninfected or E. chaffeensis -infected THP-1 cells at 2 d p.i. were lysed in modified lysis buffer (25 mM Tris, pH 7.4, 150 mM NaCl, 1% NP40 [USB, 19638], 5% glycerol, and 1% protease inhibitor cocktail III [Calbiochem, 539134]) for 15 min and immunoprecipitated for 2 h with rabbit anti-Etf-1, -BECN1, or -PIK3C3, or mouse anti-RAB5 IgG that was cross-linked to protein A/G-magnetic beads (Pierce, 88805).

    Techniques: Immunoprecipitation, Infection, Modification, Lysis, Magnetic Beads, Western Blot, Activated Clotting Time Assay

    Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus increases neutrophil transepithelial migration into the P. aeruginosa infected airspace of the lung. FACS analysis using Ly6G and CD11b markers to isolate the neutrophil population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood (PB), data are shown as mean ± SD, and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection, FACS

    Helminth-induced increase in neutrophil migration during P. aeruginosa infection is correlated with increased MPO and ELA2 levels within the airspace. Protein extracts were prepared from BAL samples and analyzed for ( a ) neutrophil myeloperoxidase (MPO) and ( b ) elastase (ELA2). ( c ) LDH levels were assessed in the cell-free supernatant of the BAL to assess cytotoxicity. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Helminth-induced increase in neutrophil migration during P. aeruginosa infection is correlated with increased MPO and ELA2 levels within the airspace. Protein extracts were prepared from BAL samples and analyzed for ( a ) neutrophil myeloperoxidase (MPO) and ( b ) elastase (ELA2). ( c ) LDH levels were assessed in the cell-free supernatant of the BAL to assess cytotoxicity. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection

    Coinfection with H. polygyrus does not impact macrophage levels within the airspace, lung tissue or circulation during P. aeruginosa infection of the airspace. FACS analysis using F4/80 and CD11b markers to isolate the macrophage populations in the ( a ) BAL, ( b ) lung tissue and ( c ) monocytes in peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus does not impact macrophage levels within the airspace, lung tissue or circulation during P. aeruginosa infection of the airspace. FACS analysis using F4/80 and CD11b markers to isolate the macrophage populations in the ( a ) BAL, ( b ) lung tissue and ( c ) monocytes in peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Infection, FACS

    Coinfection with H. polygyrus increases cellular transepithelial migration into the lung airspace. After 2 weeks of helminth infection, the mice were infected with/without P. aeruginosa through intranasal inoculation. Mice were placed in the cage for 6 hours, and after incubation mice were sacrificed, BAL and lung tissue were obtained. ( a ) The result of BAL cell count. Data are shown as mean ± SD and are representative of three independent experiments. ** P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus increases cellular transepithelial migration into the lung airspace. After 2 weeks of helminth infection, the mice were infected with/without P. aeruginosa through intranasal inoculation. Mice were placed in the cage for 6 hours, and after incubation mice were sacrificed, BAL and lung tissue were obtained. ( a ) The result of BAL cell count. Data are shown as mean ± SD and are representative of three independent experiments. ** P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: Migration, Infection, Mouse Assay, Incubation, Cell Counting

    Coinfection with H. polygyrus during P. aeruginosa acute pneumonia increases the CD4 + T-cells population in the airspace and lung tissues. FACS analysis using CD4 marker to isolate the CD4 + T-cell population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Journal: Scientific Reports

    Article Title: Intestinal helminth infection enhances bacteria-induced recruitment of neutrophils to the airspace

    doi: 10.1038/s41598-019-51991-3

    Figure Lengend Snippet: Coinfection with H. polygyrus during P. aeruginosa acute pneumonia increases the CD4 + T-cells population in the airspace and lung tissues. FACS analysis using CD4 marker to isolate the CD4 + T-cell population in the ( a ) BAL, ( b ) lung tissue and ( c ) peripheral blood. Data are shown as mean ± SD and are representative of three independent experiments. * P

    Article Snippet: For protein lysate samples, one of the 0.5 mL BAL aliquots was treated with 10 μL protease inhibitor cocktail set III, EDTA-free (Calbiochem, MA) and 20 μl of 10% Triton X-100 (Sigma, MA) and mixed by inversion for 20 minutes at 4 °C.

    Techniques: FACS, Marker

    Western blot of sLOX-1 released from cells treated with MβCD and different protease inhibitors. LOX-1-V5-COS transfected cells were incubated with MβCD, GM6001, PMSF, Pepstatin A and the protease inhibitor cocktail (PI mix) (Calbiochem), as indicated (upper panel). Histogram shows the densitometric analysis expressed as percentage of reduction of sLOX-1 released in the presence of different protease inhibitors. 100% refers as sLOX-1 released in the presence of 5mM MβCD for 30 min (lower panel). Data in histograms represent the average ± SEM of three experiments, P

    Journal: PLoS ONE

    Article Title: Membrane Cholesterol Modulates LOX-1 Shedding in Endothelial Cells

    doi: 10.1371/journal.pone.0141270

    Figure Lengend Snippet: Western blot of sLOX-1 released from cells treated with MβCD and different protease inhibitors. LOX-1-V5-COS transfected cells were incubated with MβCD, GM6001, PMSF, Pepstatin A and the protease inhibitor cocktail (PI mix) (Calbiochem), as indicated (upper panel). Histogram shows the densitometric analysis expressed as percentage of reduction of sLOX-1 released in the presence of different protease inhibitors. 100% refers as sLOX-1 released in the presence of 5mM MβCD for 30 min (lower panel). Data in histograms represent the average ± SEM of three experiments, P

    Article Snippet: Phenylmethylsulfonylfuoride (PMSF) (Euroclone, Devon, UK), pepstatin A (Boehringer, Mannheim, DE), GM6001 inhibitor (Biomol Research Lab Inc.), protease inhibitor cocktail set III (AEBSF 100mM; aprotinin 80μM; bestatin 5mM; E-64 1,5mM; leupeptin 2mM; pepstatin A 1mM) (Calbiochem, La Jolla, CA) were used for specific protease class inhibition experiments.

    Techniques: Western Blot, Transfection, Incubation, Protease Inhibitor