protease inhibitor cocktail  (thermo fisher)


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    Name:
    Protease Inhibitor Cocktails
    Description:

    Catalog Number:
    78410
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    Structured Review

    thermo fisher protease inhibitor cocktail

    https://www.bioz.com/result/protease inhibitor cocktail/product/thermo fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protease inhibitor cocktail - by Bioz Stars, 2021-03
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    Protease Inhibitor:

    Article Title: HEXIM1-Tat chimera inhibits HIV-1 replication
    Article Snippet: Transfection amounts were as follow: lane 1, 500 ng/mL pHT1 and 500ng mL pTat; lane 3, 500 ng/mL pHT1; lane 4, 500 ng/mL pTat; lanes 5–7: 500 ng/mL pTat and 250, 500 or 1,000 ng/mL pHT1; in all lanes, pEF-Bos qsp 1.5 μg/mL total DNA. .. After 48 hrs, the cells were washed using PBS, and lysed using RIPA buffer containing 150 mM KCl and a protease inhibitor mixture (Thermo Fisher Scientific). .. Lysates were precleared for 2 hrs at 4°C using protein G-sepharose beads (Life Technologyies, and the precleared lysates were incubated overnight at 4°C with 4 μg of the indicated primary Ab or control mouse IgG1 (MI10-102, Bethyl).

    Article Title: Multiplexed CRISPR/Cas9-mediated knockout of 19 Fanconi anemia pathway genes in zebrafish revealed their roles in growth, sexual development and fertility
    Article Snippet: For Tp53 samples, about twenty-five embryos collected at 24 hpf were dechorionated, deyolked and homogenized in RIPA buffer containing protease inhibitor cocktail (Thermo Fisher Scientific). .. For Fancd2 samples, soft tissue such as heart, liver, kidney and testis from adult fish were homogenized using TissueRuptor (Qiagen) in cell lysis buffer (Cell Signaling Technology) containing protease inhibitor cocktail (Thermo Fisher Scientific). ..

    Article Title: P2Y1 receptor blockade normalizes network dysfunction and cognition in an Alzheimer’s disease model
    Article Snippet: .. Protein extraction was performed by homogenization in PBS, pH 7.4, and 1% phosphatase inhibitor cocktail (Thermo Fisher) and 1% protease inhibitor cocktail (Thermo Fisher) using a ceramic bead homogenizer (Precellys; VWR). ..

    Article Title: Simple In Vitro Assay To Evaluate the Incorporation Efficiency of Ribonucleotide Analog 5′-Triphosphates into RNA by Human Mitochondrial DNA-Dependent RNA Polymerase
    Article Snippet: Urea, taurine, dithiothreitol (DTT), imidazole, MgCl2 , and IPTG (isopropyl-β- d -thiogalactopyranoside) were purchased from Sigma (St. Louis, MO). .. LB medium, bovine serum albumin (BSA), NaCl, Triton X-100, glycerol, protease inhibitor cocktail, Tris-HCl (pH 7.5), and HisPur Ni-NTA agarose resin were purchased from Thermo Fisher Scientific (Waltham, MA). .. An In-Fusion HD cloning kit and Stellar competent cells were purchased from Clontech (Mountain View, CA).

    Article Title: Human wild-type superoxide dismutase 1 gene delivery to rat bone marrow stromal cells: its importance and potential future trends
    Article Snippet: The PCR amplifications were performed using the 2X PCR master mix (Fermentas, Lithuania) with 0.5 μl of reverse transcription product for each sample of human SOD1 with rat glyceraldehyde-3-phosphate dehydrogenase ( GAPDH , housekeeping gene) and SOD1 primers which have been listed in . .. Western blot The supernatant of the transfected cells was collected on ice, and protease inhibitor cocktail (Fermentas, Lithuania) was added immediately to the samples. ..

    Fluorescence In Situ Hybridization:

    Article Title: Multiplexed CRISPR/Cas9-mediated knockout of 19 Fanconi anemia pathway genes in zebrafish revealed their roles in growth, sexual development and fertility
    Article Snippet: For Tp53 samples, about twenty-five embryos collected at 24 hpf were dechorionated, deyolked and homogenized in RIPA buffer containing protease inhibitor cocktail (Thermo Fisher Scientific). .. For Fancd2 samples, soft tissue such as heart, liver, kidney and testis from adult fish were homogenized using TissueRuptor (Qiagen) in cell lysis buffer (Cell Signaling Technology) containing protease inhibitor cocktail (Thermo Fisher Scientific). ..

    Lysis:

    Article Title: Multiplexed CRISPR/Cas9-mediated knockout of 19 Fanconi anemia pathway genes in zebrafish revealed their roles in growth, sexual development and fertility
    Article Snippet: For Tp53 samples, about twenty-five embryos collected at 24 hpf were dechorionated, deyolked and homogenized in RIPA buffer containing protease inhibitor cocktail (Thermo Fisher Scientific). .. For Fancd2 samples, soft tissue such as heart, liver, kidney and testis from adult fish were homogenized using TissueRuptor (Qiagen) in cell lysis buffer (Cell Signaling Technology) containing protease inhibitor cocktail (Thermo Fisher Scientific). ..

    Protein Extraction:

    Article Title: P2Y1 receptor blockade normalizes network dysfunction and cognition in an Alzheimer’s disease model
    Article Snippet: .. Protein extraction was performed by homogenization in PBS, pH 7.4, and 1% phosphatase inhibitor cocktail (Thermo Fisher) and 1% protease inhibitor cocktail (Thermo Fisher) using a ceramic bead homogenizer (Precellys; VWR). ..

    Homogenization:

    Article Title: P2Y1 receptor blockade normalizes network dysfunction and cognition in an Alzheimer’s disease model
    Article Snippet: .. Protein extraction was performed by homogenization in PBS, pH 7.4, and 1% phosphatase inhibitor cocktail (Thermo Fisher) and 1% protease inhibitor cocktail (Thermo Fisher) using a ceramic bead homogenizer (Precellys; VWR). ..

    Western Blot:

    Article Title: Human wild-type superoxide dismutase 1 gene delivery to rat bone marrow stromal cells: its importance and potential future trends
    Article Snippet: The PCR amplifications were performed using the 2X PCR master mix (Fermentas, Lithuania) with 0.5 μl of reverse transcription product for each sample of human SOD1 with rat glyceraldehyde-3-phosphate dehydrogenase ( GAPDH , housekeeping gene) and SOD1 primers which have been listed in . .. Western blot The supernatant of the transfected cells was collected on ice, and protease inhibitor cocktail (Fermentas, Lithuania) was added immediately to the samples. ..

    Transfection:

    Article Title: Human wild-type superoxide dismutase 1 gene delivery to rat bone marrow stromal cells: its importance and potential future trends
    Article Snippet: The PCR amplifications were performed using the 2X PCR master mix (Fermentas, Lithuania) with 0.5 μl of reverse transcription product for each sample of human SOD1 with rat glyceraldehyde-3-phosphate dehydrogenase ( GAPDH , housekeeping gene) and SOD1 primers which have been listed in . .. Western blot The supernatant of the transfected cells was collected on ice, and protease inhibitor cocktail (Fermentas, Lithuania) was added immediately to the samples. ..

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  • 97
    Thermo Fisher halt protease inhibitor cocktail
    Peptidase activity assays of purified recombinant FliA(H)-hypervariable region and flagellar filaments from C. haemolyticum . a 12% SDS-PAGE of purified FliA(H)-hypervariable region (band “i”), electrophoresing close to that predicted for recombinant thioredoxin-tagged flagellin hypervariable region (50.14 kDa). b Amino acid occurrence heat map based on FliA(H)-hypervariable region protease cleaved peptide alignments of 391 cleavage sites identified in a tryptic E. coli K12 library. c IceLogos of FliA(H)-hypervariable region protease cleaved peptide specificity profiles showing percent differences compared to natural amino acid abundance, with significantly over-represented amino acids shown above the x -axis and under-represented residues below the x -axis. Amino acids that have not been identified are depicted in pink . d Fluorometric peptidase assays for FliA(H)-hypervariable region protease against three different peptidic substrates (ALG↓L, PLG↓L, PLG↓V) and + /– <t>EDTA</t> as indicated. e 10% SDS-PAGE of purified flagellar sheared filaments from C. haemolyticum . Band “ii” and “iii” were identified by mass spectrometry as FliA(H) (proteolytic flagellin) and the non-protease-containing structural flagellin (WP_039229459), respectively (Supplementary Fig. 7 ). f Amino acid occurrence heat map and g IceLogo based on cleaved peptide alignments of 269 cleavage sites identified in a tryptic E. coli K12 library using purified flagellar filaments. h Fluorometric peptidase assays for flagellar filaments using the ALG↓L peptide in three assay conditions including EDTA-free <t>HALT</t> protease inhibitors
    Halt Protease Inhibitor Cocktail, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/halt protease inhibitor cocktail/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
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    97
    Thermo Fisher 1x ethylene diamine triacetic acid edta
    Lentiviral vector is released from LDLR3-ELP after internalization by cells A549 were starved for 24 hours then were treated in serum free media with the indicated treatments. Cells were treated for 24 hours with the lentiviral vector containing nanoparticles that consisted of 1 µM of biotinylated LDLR3-ELP and 1 µM KGF-ELP. At the indicated time points, cells were put on ice for five minutes followed by three ice cold washed in PBS. The cells were lysed with radio immune protection assay (RIPA) buffer containing 2X halt protease and phosphatase inhibitors cocktail with <t>1X</t> ethylene diamine <t>triacetic</t> acid <t>(EDTA).</t> The lysates were immunoprecipitated using streptavidin conjugated magnetic beads (dynabeads). Then a magnetic bar was used to separate the beads from the supernatant. Following a few washes, the immunoprecipitated complexes (dynabeads) and the supernatant were analyzed for the presence of virus using western blots. Anti VSV-G antibody was used to detect the lentiviral vector.
    1x Ethylene Diamine Triacetic Acid Edta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x ethylene diamine triacetic acid edta/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
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    99
    Thermo Fisher 1x halt protease
    Analysis of protein expression. Cell-seeded scaffolds were randomly collected, trypsinized, lysed with IP lysis buffer—1X Halt protease and phosphatase inhibitor cocktail and cell lysates were subjected to SDS-PAGE followed by western blotting. (a) In separate experiments, the membranes were incubated with primary antibodies against ACAN, COL1A1, COL2A2, COL9A1, SOX-9, p-SOX-9 (i.e. p-S181-SOX-9). β-Actin was used as the loading control. (b) All protein bands were quantified by ImageJ ™  software and relative protein expression levels were computed with respect to β-actin.
    1x Halt Protease, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x halt protease/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1x halt protease - by Bioz Stars, 2021-03
    99/100 stars
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    Image Search Results


    Peptidase activity assays of purified recombinant FliA(H)-hypervariable region and flagellar filaments from C. haemolyticum . a 12% SDS-PAGE of purified FliA(H)-hypervariable region (band “i”), electrophoresing close to that predicted for recombinant thioredoxin-tagged flagellin hypervariable region (50.14 kDa). b Amino acid occurrence heat map based on FliA(H)-hypervariable region protease cleaved peptide alignments of 391 cleavage sites identified in a tryptic E. coli K12 library. c IceLogos of FliA(H)-hypervariable region protease cleaved peptide specificity profiles showing percent differences compared to natural amino acid abundance, with significantly over-represented amino acids shown above the x -axis and under-represented residues below the x -axis. Amino acids that have not been identified are depicted in pink . d Fluorometric peptidase assays for FliA(H)-hypervariable region protease against three different peptidic substrates (ALG↓L, PLG↓L, PLG↓V) and + /– EDTA as indicated. e 10% SDS-PAGE of purified flagellar sheared filaments from C. haemolyticum . Band “ii” and “iii” were identified by mass spectrometry as FliA(H) (proteolytic flagellin) and the non-protease-containing structural flagellin (WP_039229459), respectively (Supplementary Fig. 7 ). f Amino acid occurrence heat map and g IceLogo based on cleaved peptide alignments of 269 cleavage sites identified in a tryptic E. coli K12 library using purified flagellar filaments. h Fluorometric peptidase assays for flagellar filaments using the ALG↓L peptide in three assay conditions including EDTA-free HALT protease inhibitors

    Journal: Nature Communications

    Article Title: Discovery of a proteolytic flagellin family in diverse bacterial phyla that assembles enzymatically active flagella

    doi: 10.1038/s41467-017-00599-0

    Figure Lengend Snippet: Peptidase activity assays of purified recombinant FliA(H)-hypervariable region and flagellar filaments from C. haemolyticum . a 12% SDS-PAGE of purified FliA(H)-hypervariable region (band “i”), electrophoresing close to that predicted for recombinant thioredoxin-tagged flagellin hypervariable region (50.14 kDa). b Amino acid occurrence heat map based on FliA(H)-hypervariable region protease cleaved peptide alignments of 391 cleavage sites identified in a tryptic E. coli K12 library. c IceLogos of FliA(H)-hypervariable region protease cleaved peptide specificity profiles showing percent differences compared to natural amino acid abundance, with significantly over-represented amino acids shown above the x -axis and under-represented residues below the x -axis. Amino acids that have not been identified are depicted in pink . d Fluorometric peptidase assays for FliA(H)-hypervariable region protease against three different peptidic substrates (ALG↓L, PLG↓L, PLG↓V) and + /– EDTA as indicated. e 10% SDS-PAGE of purified flagellar sheared filaments from C. haemolyticum . Band “ii” and “iii” were identified by mass spectrometry as FliA(H) (proteolytic flagellin) and the non-protease-containing structural flagellin (WP_039229459), respectively (Supplementary Fig. 7 ). f Amino acid occurrence heat map and g IceLogo based on cleaved peptide alignments of 269 cleavage sites identified in a tryptic E. coli K12 library using purified flagellar filaments. h Fluorometric peptidase assays for flagellar filaments using the ALG↓L peptide in three assay conditions including EDTA-free HALT protease inhibitors

    Article Snippet: Purified recombinant FliA(H)-hypervariable region protease was incubated at a final concentration of 0.5 µM with 10 µM QF-peptide substrate in 100 µl of 150 mM NaCl, 10 mM CaCl2 , 50 mM HEPES, pH 7.5, in the presence of HALT protease inhibitor cocktail (Life Technologies) plus/minus 20 mM EDTA at 37 °C.

    Techniques: Activity Assay, Purification, Recombinant, SDS Page, Mass Spectrometry

    Lentiviral vector is released from LDLR3-ELP after internalization by cells A549 were starved for 24 hours then were treated in serum free media with the indicated treatments. Cells were treated for 24 hours with the lentiviral vector containing nanoparticles that consisted of 1 µM of biotinylated LDLR3-ELP and 1 µM KGF-ELP. At the indicated time points, cells were put on ice for five minutes followed by three ice cold washed in PBS. The cells were lysed with radio immune protection assay (RIPA) buffer containing 2X halt protease and phosphatase inhibitors cocktail with 1X ethylene diamine triacetic acid (EDTA). The lysates were immunoprecipitated using streptavidin conjugated magnetic beads (dynabeads). Then a magnetic bar was used to separate the beads from the supernatant. Following a few washes, the immunoprecipitated complexes (dynabeads) and the supernatant were analyzed for the presence of virus using western blots. Anti VSV-G antibody was used to detect the lentiviral vector.

    Journal: Gene therapy

    Article Title: Recombinant elastin based nanoparticles for targeted gene therapy

    doi: 10.1038/gt.2017.54

    Figure Lengend Snippet: Lentiviral vector is released from LDLR3-ELP after internalization by cells A549 were starved for 24 hours then were treated in serum free media with the indicated treatments. Cells were treated for 24 hours with the lentiviral vector containing nanoparticles that consisted of 1 µM of biotinylated LDLR3-ELP and 1 µM KGF-ELP. At the indicated time points, cells were put on ice for five minutes followed by three ice cold washed in PBS. The cells were lysed with radio immune protection assay (RIPA) buffer containing 2X halt protease and phosphatase inhibitors cocktail with 1X ethylene diamine triacetic acid (EDTA). The lysates were immunoprecipitated using streptavidin conjugated magnetic beads (dynabeads). Then a magnetic bar was used to separate the beads from the supernatant. Following a few washes, the immunoprecipitated complexes (dynabeads) and the supernatant were analyzed for the presence of virus using western blots. Anti VSV-G antibody was used to detect the lentiviral vector.

    Article Snippet: The cells were then lysed with radio immune protection assay (RIPA) buffer containing 2X halt protease and phosphatase inhibitors cocktail with 1X ethylene diamine triacetic acid (EDTA) (Thermo scientific cat #78440).

    Techniques: Plasmid Preparation, Immunoprecipitation, Magnetic Beads, Western Blot

    N-Cadherin from POMGNT1-deficient cells shows enhanced homotypic interactions in vitro . (A) Semi-quantitative analysis of the mean size of dynabead aggregates coated with the recombinant extracellular domain of recombinant N-Cdh purified from WT (gray line) and ΔPOMGnT1 cells (black line). Protein-coated dynabeads were imaged following incubation in the presence of either CaCl 2 (solid lines) or EDTA (broken lines) at the indicated time intervals. Mean aggregate size is plotted against time. Assays were performed at quadruplicate from two independent experiments. Data are represented as means ± SD. Asterisks denote statistical significance in comparison to WT cells: * p ≤ 0.05, ** p ≤ 0.01. (B) Western Blot analysis evaluating levels of protein from WT and ΔPOMGnT1 (Δ) bound to dynabeads. Dynabead-bound recombinant N-Cdh was eluted in SDS sample buffer (beads), resolved on a 10% SDS PAA gel. For detection on Western blot an anti-5xHis antibody was used. In addition, protein before binding to beads (input) and unbound fractions (void) are shown.

    Journal: bioRxiv

    Article Title: Glycosyltransferase POMGNT1 deficiency affects N-cadherin-mediated cell-cell adhesion

    doi: 10.1101/2020.09.09.289306

    Figure Lengend Snippet: N-Cadherin from POMGNT1-deficient cells shows enhanced homotypic interactions in vitro . (A) Semi-quantitative analysis of the mean size of dynabead aggregates coated with the recombinant extracellular domain of recombinant N-Cdh purified from WT (gray line) and ΔPOMGnT1 cells (black line). Protein-coated dynabeads were imaged following incubation in the presence of either CaCl 2 (solid lines) or EDTA (broken lines) at the indicated time intervals. Mean aggregate size is plotted against time. Assays were performed at quadruplicate from two independent experiments. Data are represented as means ± SD. Asterisks denote statistical significance in comparison to WT cells: * p ≤ 0.05, ** p ≤ 0.01. (B) Western Blot analysis evaluating levels of protein from WT and ΔPOMGnT1 (Δ) bound to dynabeads. Dynabead-bound recombinant N-Cdh was eluted in SDS sample buffer (beads), resolved on a 10% SDS PAA gel. For detection on Western blot an anti-5xHis antibody was used. In addition, protein before binding to beads (input) and unbound fractions (void) are shown.

    Article Snippet: Whole cell lysate and crude membrane preparation Cells at 90% confluency (~5.0 x 106 cells) were washed with ice-cold PBS and lysed with 500 μl of lysis buffer (10 mM Tris-HCl pH 7.6, 1% (w/v) SDS, 1 mM EDTA, supplemented with Halt™ protease and phosphatase inhibitor cocktail without EDTA (ThermoFisher Scientific)).

    Techniques: In Vitro, Recombinant, Purification, Incubation, Western Blot, Binding Assay

    Analysis of protein expression. Cell-seeded scaffolds were randomly collected, trypsinized, lysed with IP lysis buffer—1X Halt protease and phosphatase inhibitor cocktail and cell lysates were subjected to SDS-PAGE followed by western blotting. (a) In separate experiments, the membranes were incubated with primary antibodies against ACAN, COL1A1, COL2A2, COL9A1, SOX-9, p-SOX-9 (i.e. p-S181-SOX-9). β-Actin was used as the loading control. (b) All protein bands were quantified by ImageJ ™  software and relative protein expression levels were computed with respect to β-actin.

    Journal: Journal of Tissue Engineering

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis

    doi: 10.1177/2041731414566529

    Figure Lengend Snippet: Analysis of protein expression. Cell-seeded scaffolds were randomly collected, trypsinized, lysed with IP lysis buffer—1X Halt protease and phosphatase inhibitor cocktail and cell lysates were subjected to SDS-PAGE followed by western blotting. (a) In separate experiments, the membranes were incubated with primary antibodies against ACAN, COL1A1, COL2A2, COL9A1, SOX-9, p-SOX-9 (i.e. p-S181-SOX-9). β-Actin was used as the loading control. (b) All protein bands were quantified by ImageJ ™ software and relative protein expression levels were computed with respect to β-actin.

    Article Snippet: Protein isolation and western blotting analysis At the end of 14 days of culture, cells were released from scaffolds with 1X Trypsin followed by pelleting cells at 1000×g and finally lysing the pellet with Pierce IP lysis buffer supplemented with 1X Halt protease and phosphatase inhibitor cocktail (Thermo Scientific, Rockford, IL, USA).

    Techniques: Expressing, Lysis, SDS Page, Western Blot, Incubation, Software