phosphatase inhibitor cocktail  (Thermo Fisher)


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    Name:
    Halt Phosphatase Inhibitor Cocktail
    Description:
    Thermo Scientific Halt Phosphatase Inhibitor Cocktail preserves the phosphorylation state of proteins during and after cell lysis or tissue protein extraction Features of Halt Phosphatase Inhibitor Cocktail • Devoid of protease inhibitors To protect against both proteases and phosphatases use Halt Protease and Phosphatase Inhibitor Cocktail • Dialyzable Either dialyze or desalt sample to effectively remove inhibitors from cell extracts before performing procedures that are discovered to be incompatible with cocktail components • Supplied as a 100X concentrate effective when used at a final concentration of 1X however samples that contain particularly high levels of phosphatase activity may require a final concentration of 2 3X • Compatibility compatible with most detergent based cell lysis reagents including all Thermo Scientific Pierce Cell Lysis Solutions • All in one format inhibits both serine threonine phosphatases and protein tyrosine phosphatases • Compatiblewith popular protein assays works with standard protein assays including BCA and coomassie Bradford assays • Convenient storage refrigerator 2 to 8°C storage saves valuable space in the freezer and provides for immediate use • Multiple sizes select a package size of the 100X cocktail that is ideal for any particular scale and workflow of protein extraction experiment Halt Phosphatase Inhibitor Cocktail protects phosphoproteins from serine threonine phosphatases as well as protein tyrosine phosphatases PTPs during cell lysis The cocktail contains a mixture of four inhibitors of broad specificity including sodium fluoride sodium orthovanadate sodium pyrophosphate and beta glycerophosphate Conveniently packaged in a 100X format the cocktail is stable at 4°C and is suitable for use with any cell or tissue extract The cocktail preserves protein phosphorylation in immunoprecipitation and kinase assays Phosphorylation and dephosphorylation comprise a molecular on off switch that regulates many key biological pathways within the cell including signal transduction cell division and apoptosis To effectively study phosphorylation activation states one must inactivate phosphatases to prevent their uncontrolled activity upon cell lysis Halt Phosphatase Inhibitor Cocktail accomplishes this task Related Products Pierce Phosphatase Inhibitor Mini Tablets
    Catalog Number:
    78420
    Price:
    None
    Category:
    Lab Reagents and Chemicals
    Applications:
    Cell Lysis & Fractionation|Protease and Phosphatase Inhibition|Protein Biology|Protein Purification & Isolation
    Buy from Supplier


    Structured Review

    Thermo Fisher phosphatase inhibitor cocktail
    Thermo Scientific Halt Phosphatase Inhibitor Cocktail preserves the phosphorylation state of proteins during and after cell lysis or tissue protein extraction Features of Halt Phosphatase Inhibitor Cocktail • Devoid of protease inhibitors To protect against both proteases and phosphatases use Halt Protease and Phosphatase Inhibitor Cocktail • Dialyzable Either dialyze or desalt sample to effectively remove inhibitors from cell extracts before performing procedures that are discovered to be incompatible with cocktail components • Supplied as a 100X concentrate effective when used at a final concentration of 1X however samples that contain particularly high levels of phosphatase activity may require a final concentration of 2 3X • Compatibility compatible with most detergent based cell lysis reagents including all Thermo Scientific Pierce Cell Lysis Solutions • All in one format inhibits both serine threonine phosphatases and protein tyrosine phosphatases • Compatiblewith popular protein assays works with standard protein assays including BCA and coomassie Bradford assays • Convenient storage refrigerator 2 to 8°C storage saves valuable space in the freezer and provides for immediate use • Multiple sizes select a package size of the 100X cocktail that is ideal for any particular scale and workflow of protein extraction experiment Halt Phosphatase Inhibitor Cocktail protects phosphoproteins from serine threonine phosphatases as well as protein tyrosine phosphatases PTPs during cell lysis The cocktail contains a mixture of four inhibitors of broad specificity including sodium fluoride sodium orthovanadate sodium pyrophosphate and beta glycerophosphate Conveniently packaged in a 100X format the cocktail is stable at 4°C and is suitable for use with any cell or tissue extract The cocktail preserves protein phosphorylation in immunoprecipitation and kinase assays Phosphorylation and dephosphorylation comprise a molecular on off switch that regulates many key biological pathways within the cell including signal transduction cell division and apoptosis To effectively study phosphorylation activation states one must inactivate phosphatases to prevent their uncontrolled activity upon cell lysis Halt Phosphatase Inhibitor Cocktail accomplishes this task Related Products Pierce Phosphatase Inhibitor Mini Tablets
    https://www.bioz.com/result/phosphatase inhibitor cocktail/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphatase inhibitor cocktail - by Bioz Stars, 2021-03
    99/100 stars

    Images

    Related Articles

    Isolation:

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: The amplified expression of mRNA transcripts was normalized to GAPDH expression; 2−ΔΔCt method was used to calculate relative expression levels. .. 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). .. After centrifugation of the lysate at 15,000×g for 10 min at 4°C, supernatants were collected and the protein concentration was measured by the bicinchoninic acid (BCA) method.

    Western Blot:

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: The amplified expression of mRNA transcripts was normalized to GAPDH expression; 2−ΔΔCt method was used to calculate relative expression levels. .. 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). .. After centrifugation of the lysate at 15,000×g for 10 min at 4°C, supernatants were collected and the protein concentration was measured by the bicinchoninic acid (BCA) method.

    Article Title: Impaired myogenic development, differentiation and function in hESC-derived SMA myoblasts and myotubes
    Article Snippet: Stained membranes were scanned with an Odyssey Infrared Imager (#9120, Li-Cor) and values normalized to internal control. .. For capillary western blotting, cells were lysed for 5 min using 1x RIPA buffer (#R0278, Sigma-Aldrich) with Phosphatase Inhibitor Cocktail (#04906845001, Roche Applied Science) and complete Protease Inhibitor Cocktail (#04693124001, Roche Applied Science) or RIPA supplemented with Halt Protease inhibitor cocktail (#20–188, Millipore) and Halt Phosphatase inhibitor cocktail (#1861277, ThermoFisher Scientific). .. Total protein in the supernatant was quantified by bicinchoninic acid assay (#23225, ThermoFisher Scientific) or Nanodrop 1000.

    Lysis:

    Article Title: Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis
    Article Snippet: The amplified expression of mRNA transcripts was normalized to GAPDH expression; 2−ΔΔCt method was used to calculate relative expression levels. .. 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). .. After centrifugation of the lysate at 15,000×g for 10 min at 4°C, supernatants were collected and the protein concentration was measured by the bicinchoninic acid (BCA) method.

    Article Title: March1-dependent modulation of donor MHC II on CD103+ dendritic cells mitigates alloimmunity
    Article Snippet: .. Cells were then washed with ice-cold Dulbecco’s PBS and lysed in NP-40 lysis buffer (containing 1% NP-40, 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM phenylmethylsulfonyl fluoride (PMSF), 1× HALT protease and phosphatase inhibitor cocktail (Thermo Scientific)). .. For immunoprecipitation (IP), 1 mg of cell extract was incubated with 5 µg of anti-I-A/I-E (MHC II—clone M5/114, eBioscience) for 2 h at 4 °C followed by incubation with 20 µL of protein A (50% slurry, GE Healthcare) plus sepharose beads for either 2 h at room temperature or overnight at 4 °C.

    Protease Inhibitor:

    Article Title: Impaired myogenic development, differentiation and function in hESC-derived SMA myoblasts and myotubes
    Article Snippet: Stained membranes were scanned with an Odyssey Infrared Imager (#9120, Li-Cor) and values normalized to internal control. .. For capillary western blotting, cells were lysed for 5 min using 1x RIPA buffer (#R0278, Sigma-Aldrich) with Phosphatase Inhibitor Cocktail (#04906845001, Roche Applied Science) and complete Protease Inhibitor Cocktail (#04693124001, Roche Applied Science) or RIPA supplemented with Halt Protease inhibitor cocktail (#20–188, Millipore) and Halt Phosphatase inhibitor cocktail (#1861277, ThermoFisher Scientific). .. Total protein in the supernatant was quantified by bicinchoninic acid assay (#23225, ThermoFisher Scientific) or Nanodrop 1000.

    Sonication:

    Article Title: Hypoxia-induced 26S proteasome dysfunction increases immunogenicity of mesenchymal stem cells
    Article Snippet: The first dimension BN-PAGE and second dimension SDS-PAGE were performed as described previously . .. Briefly, the cell lysates were prepared by sonication in 20 mM Bis-tris, 500 mM ε-aminocaproic acid 20 mM NaCl, 2 mM EDTA (pH 8.0), and Glycerol 10% supplemented with 1 × Halt protease and phosphatase inhibitor cocktail (Thermo Scientific), and 1.5% n -Dodecyl β- d -maltoside (Sigma). ..

    Article Title: Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury
    Article Snippet: The quantification of Ezh2 protein in RGCs was assessed using protein lysates of purified RGCs as previously described [ ]. .. Briefly, RGCs purified from new born (P0) mouse pups were lysed by sonication in ice-cold RIPA buffer (Cat. No. 20–188; Milipore, Billerica, MA) containing proteinase cocktail inhibitor (Ref: 05892953001; Roche, Indianapolis, IN) and phosphatase inhibitor cocktail (Cat. No. 78420; Thermo Scientific, Waltham, MA). .. Protein concentration was measured using a NanoDrop 2000 Spectrophotometer (Thermo Scientific).

    Purification:

    Article Title: Ezh2 does not mediate retinal ganglion cell homeostasis or their susceptibility to injury
    Article Snippet: The quantification of Ezh2 protein in RGCs was assessed using protein lysates of purified RGCs as previously described [ ]. .. Briefly, RGCs purified from new born (P0) mouse pups were lysed by sonication in ice-cold RIPA buffer (Cat. No. 20–188; Milipore, Billerica, MA) containing proteinase cocktail inhibitor (Ref: 05892953001; Roche, Indianapolis, IN) and phosphatase inhibitor cocktail (Cat. No. 78420; Thermo Scientific, Waltham, MA). .. Protein concentration was measured using a NanoDrop 2000 Spectrophotometer (Thermo Scientific).

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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
    Price from $9.99 to $1999.99
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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
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    97
    Thermo Fisher halt protease and phosphatase inhibitor single use cocktail 100x
    4-HNE accumulates during intracellular bacterial infection by L. monocytogenes . (A) 4-HNE accumulation in TIB73 murine hepatocytes during intracellular L. monocytogenes infection. 4-HNE adduct accumulation is assessed by dot blot whole cell Western and normalized to actin levels. Data are normalized 4-HNE levels as percent of 4-HNE at hour 0 of infection. Western image below is representative. (B) 4-HNE accumulation during 48-hour murine infection by GFP + L. monocytogenes assessed by immunohistochemistry analysis. Spleen (infected) anti-4-HNE. (C) spleen (uninfected) anti-4-HNE. (D) spleen (infected) anti-GFP 25x magnification. (E) spleen (infected) anti-4-HNE 25x magnification (F) spleen (infected) anti-GFP <t>100x</t> magnification. (G) spleen (infected) anti-4-HNE 100x magnification. Antigens detected with 3,3-diaminobenzidine staining by horseradish peroxidase and cellular nuclei imaged with Hematoxylin counterstain. Data in (A) are pooled from two independent experiments. Statistics in (A) are an ordinary one-way ANOVA with a Dunnett’s multiple comparison test against hour 0. Error bars are mean +/- SD. *, p
    Halt Protease And Phosphatase Inhibitor Single Use Cocktail 100x, 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 and phosphatase inhibitor single use cocktail 100x/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    halt protease and phosphatase inhibitor single use cocktail 100x - by Bioz Stars, 2021-03
    97/100 stars
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    Image Search Results


    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

    4-HNE accumulates during intracellular bacterial infection by L. monocytogenes . (A) 4-HNE accumulation in TIB73 murine hepatocytes during intracellular L. monocytogenes infection. 4-HNE adduct accumulation is assessed by dot blot whole cell Western and normalized to actin levels. Data are normalized 4-HNE levels as percent of 4-HNE at hour 0 of infection. Western image below is representative. (B) 4-HNE accumulation during 48-hour murine infection by GFP + L. monocytogenes assessed by immunohistochemistry analysis. Spleen (infected) anti-4-HNE. (C) spleen (uninfected) anti-4-HNE. (D) spleen (infected) anti-GFP 25x magnification. (E) spleen (infected) anti-4-HNE 25x magnification (F) spleen (infected) anti-GFP 100x magnification. (G) spleen (infected) anti-4-HNE 100x magnification. Antigens detected with 3,3-diaminobenzidine staining by horseradish peroxidase and cellular nuclei imaged with Hematoxylin counterstain. Data in (A) are pooled from two independent experiments. Statistics in (A) are an ordinary one-way ANOVA with a Dunnett’s multiple comparison test against hour 0. Error bars are mean +/- SD. *, p

    Journal: bioRxiv

    Article Title: 4-hydroxy-2-nonenal antimicrobial toxicity is neutralized by an intracellular pathogen

    doi: 10.1101/2020.05.25.115097

    Figure Lengend Snippet: 4-HNE accumulates during intracellular bacterial infection by L. monocytogenes . (A) 4-HNE accumulation in TIB73 murine hepatocytes during intracellular L. monocytogenes infection. 4-HNE adduct accumulation is assessed by dot blot whole cell Western and normalized to actin levels. Data are normalized 4-HNE levels as percent of 4-HNE at hour 0 of infection. Western image below is representative. (B) 4-HNE accumulation during 48-hour murine infection by GFP + L. monocytogenes assessed by immunohistochemistry analysis. Spleen (infected) anti-4-HNE. (C) spleen (uninfected) anti-4-HNE. (D) spleen (infected) anti-GFP 25x magnification. (E) spleen (infected) anti-4-HNE 25x magnification (F) spleen (infected) anti-GFP 100x magnification. (G) spleen (infected) anti-4-HNE 100x magnification. Antigens detected with 3,3-diaminobenzidine staining by horseradish peroxidase and cellular nuclei imaged with Hematoxylin counterstain. Data in (A) are pooled from two independent experiments. Statistics in (A) are an ordinary one-way ANOVA with a Dunnett’s multiple comparison test against hour 0. Error bars are mean +/- SD. *, p

    Article Snippet: Bacterial infection dot blotTIB73 cells were lysed in whole cell lysis buffer (50 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X-100) with EDTA (1 µM) and Halt Protease Inhibitor Cocktail (Thermo Fisher Scientific, #78442).

    Techniques: Infection, Dot Blot, Western Blot, Immunohistochemistry, Staining