phosphatase inhibitor  (Thermo Fisher)


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    Name:
    Halt Protease and Phosphatase Inhibitor Single Use Cocktail 100X
    Description:
    Thermo Scientific Halt Protease and Phosphatase Inhibitor Single Use Cocktail 100X provides the convenience of a single solution with full protein sample protection for cell and tissue lysates Features of Halt Protease and Phosphatase Inhibitor Single Use Cocktail 100X • Complete protection all in one cocktail contains both protease and phosphatase inhibitors • Multiple package sizes choose from four convenient packages sizes to match nearly any scale • Convenient the 100X liquid formulation is much easier and more effective to use than individual inhibitors and other formats • Easy storage aqueous based format allows for convenient cold room storage without freezing by contrast with DMSO based cocktails • Compatible can be used in mass spectrometry MS because it does not contain AEBSF which can cause peaks to shift • Validated fully compatible with Thermo Scientific Pierce Cell Lysis Buffers Halt Protease and Phosphatase Inhibitor Cocktail is specifically optimized to protect proteins from degradation during extraction and purification The cocktail contains inhibitors against the major classes of proteases and phosphatases targeting aminopeptidases cysteine and serine proteases and serine threonine and protein tyrosine phosphatases To promote the inhibition of metalloproteases 0 5M EDTA is provided in a separate tube Related Products Halt Protease and Phosphatase Inhibitor Cocktail EDTA free 100X
    Catalog Number:
    78442
    Price:
    None
    Category:
    Lab Reagents and Chemicals
    Applications:
    Cell Lysis & Fractionation|Protease and Phosphatase Inhibition|Protein Biology|Protein Purification & Isolation
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    Structured Review

    Thermo Fisher phosphatase inhibitor
    Thermo Scientific Halt Protease and Phosphatase Inhibitor Single Use Cocktail 100X provides the convenience of a single solution with full protein sample protection for cell and tissue lysates Features of Halt Protease and Phosphatase Inhibitor Single Use Cocktail 100X • Complete protection all in one cocktail contains both protease and phosphatase inhibitors • Multiple package sizes choose from four convenient packages sizes to match nearly any scale • Convenient the 100X liquid formulation is much easier and more effective to use than individual inhibitors and other formats • Easy storage aqueous based format allows for convenient cold room storage without freezing by contrast with DMSO based cocktails • Compatible can be used in mass spectrometry MS because it does not contain AEBSF which can cause peaks to shift • Validated fully compatible with Thermo Scientific Pierce Cell Lysis Buffers Halt Protease and Phosphatase Inhibitor Cocktail is specifically optimized to protect proteins from degradation during extraction and purification The cocktail contains inhibitors against the major classes of proteases and phosphatases targeting aminopeptidases cysteine and serine proteases and serine threonine and protein tyrosine phosphatases To promote the inhibition of metalloproteases 0 5M EDTA is provided in a separate tube Related Products Halt Protease and Phosphatase Inhibitor Cocktail EDTA free 100X
    https://www.bioz.com/result/phosphatase inhibitor/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphatase inhibitor - by Bioz Stars, 2021-04
    97/100 stars

    Images

    Related Articles

    Infection:

    Article Title: 4-hydroxy-2-nonenal antimicrobial toxicity is neutralized by an intracellular pathogen
    Article Snippet: DNA sequencing was performed by Genewiz Incorporated. .. 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). .. Protein concentration was determined using the Pierce BCA protein assay kit (Fisher Scientific, #PI23227).

    Lysis:

    Article Title: 4-hydroxy-2-nonenal antimicrobial toxicity is neutralized by an intracellular pathogen
    Article Snippet: DNA sequencing was performed by Genewiz Incorporated. .. 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). .. Protein concentration was determined using the Pierce BCA protein assay kit (Fisher Scientific, #PI23227).

    Protease Inhibitor:

    Article Title: 4-hydroxy-2-nonenal antimicrobial toxicity is neutralized by an intracellular pathogen
    Article Snippet: DNA sequencing was performed by Genewiz Incorporated. .. 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). .. Protein concentration was determined using the Pierce BCA protein assay kit (Fisher Scientific, #PI23227).

    other:

    Article Title: 4-hydroxy-2-nonenal antimicrobial toxicity is neutralized by an intracellular pathogen
    Article Snippet: DNA sequencing was performed by Genewiz Incorporated.

    Western Blot:

    Article Title: Mechanisms of nuclear content loading to exosomes
    Article Snippet: Images were acquired using a Zeiss LSM 710 confocal microscope with a 63× objective. .. Western blotting For Western blotting, harvested cells and exosomes were lysed with radioimmunoprecipitation assay buffer (RIPA) [25 mM tris (pH 7.5), 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton X-100] supplemented with single-use phosphatase and protease inhibitors (78442, Thermo Fisher Scientific). .. Protein concentration was determined using the Micro BCA Protein Assay Kit (23235, Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Radio Immunoprecipitation:

    Article Title: Mechanisms of nuclear content loading to exosomes
    Article Snippet: Images were acquired using a Zeiss LSM 710 confocal microscope with a 63× objective. .. Western blotting For Western blotting, harvested cells and exosomes were lysed with radioimmunoprecipitation assay buffer (RIPA) [25 mM tris (pH 7.5), 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton X-100] supplemented with single-use phosphatase and protease inhibitors (78442, Thermo Fisher Scientific). .. Protein concentration was determined using the Micro BCA Protein Assay Kit (23235, Thermo Fisher Scientific) according to the manufacturer’s protocol.

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  • 85
    Thermo Fisher gene exp ppp1r1b hs00938414 m1
    Neuroimaging analyses (structure and function) of common haplotype in <t>PPP1R1B</t> in control sample of white subjects. Top row shows haplotype effects on volume ( A ) or activation ( C and E ) in striatum; bottom row shows haplotype effects on structural ( B ) and functional ( D and F ) connectivity of striatum with prefrontal cortex. Structural MRI analyses (voxel-based morphometry): ( A ) significantly reduced volume in striatum ( P
    Gene Exp Ppp1r1b Hs00938414 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gene exp ppp1r1b hs00938414 m1/product/Thermo Fisher
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    85/100 stars
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    97
    Thermo Fisher sheep spinophilin antibody
    Generation and characterization of Cre-expressing, HA-tagged human <t>spinophilin</t> mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.
    Sheep Spinophilin Antibody, 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/sheep spinophilin antibody/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sheep spinophilin antibody - by Bioz Stars, 2021-04
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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, 2021-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Neuroimaging analyses (structure and function) of common haplotype in PPP1R1B in control sample of white subjects. Top row shows haplotype effects on volume ( A ) or activation ( C and E ) in striatum; bottom row shows haplotype effects on structural ( B ) and functional ( D and F ) connectivity of striatum with prefrontal cortex. Structural MRI analyses (voxel-based morphometry): ( A ) significantly reduced volume in striatum ( P

    Journal: Journal of Clinical Investigation

    Article Title: Genetic evidence implicating DARPP-32 in human frontostriatal structure, function, and cognition

    doi: 10.1172/JCI30413

    Figure Lengend Snippet: Neuroimaging analyses (structure and function) of common haplotype in PPP1R1B in control sample of white subjects. Top row shows haplotype effects on volume ( A ) or activation ( C and E ) in striatum; bottom row shows haplotype effects on structural ( B ) and functional ( D and F ) connectivity of striatum with prefrontal cortex. Structural MRI analyses (voxel-based morphometry): ( A ) significantly reduced volume in striatum ( P

    Article Snippet: Three probes were based on the common exon structure as found in full-length cDNA: exons 4 and 5 (type 9967; Hs00259967), 1 and 2 (type 10; Hs00938410_m1) and 5 and 6 (type 14; Hs00938414_m1); splice variant 5183, exons 2 and 5; and t-DARPP (type 16; Hs00938416_g1).

    Techniques: Activation Assay, Functional Assay, Magnetic Resonance Imaging

    Genetic variation in PPP1R1B.

    Journal: Journal of Clinical Investigation

    Article Title: Genetic evidence implicating DARPP-32 in human frontostriatal structure, function, and cognition

    doi: 10.1172/JCI30413

    Figure Lengend Snippet: Genetic variation in PPP1R1B.

    Article Snippet: Three probes were based on the common exon structure as found in full-length cDNA: exons 4 and 5 (type 9967; Hs00259967), 1 and 2 (type 10; Hs00938410_m1) and 5 and 6 (type 14; Hs00938414_m1); splice variant 5183, exons 2 and 5; and t-DARPP (type 16; Hs00938416_g1).

    Techniques:

    Effect of PPP1R1B haplotype on mRNA expression in postmortem human brain. ( A ) Exon position of primers used for quantitative PCR for isoform amplification, showing which isoform was amplified by which probe. ( B ) Effect of PPP1R1B haplotypes on mRNA expression for the common isoform. The PPP1R1B haplotype has an impact on expression of all probes assaying the expression of the full-length mRNA (type 14, P

    Journal: Journal of Clinical Investigation

    Article Title: Genetic evidence implicating DARPP-32 in human frontostriatal structure, function, and cognition

    doi: 10.1172/JCI30413

    Figure Lengend Snippet: Effect of PPP1R1B haplotype on mRNA expression in postmortem human brain. ( A ) Exon position of primers used for quantitative PCR for isoform amplification, showing which isoform was amplified by which probe. ( B ) Effect of PPP1R1B haplotypes on mRNA expression for the common isoform. The PPP1R1B haplotype has an impact on expression of all probes assaying the expression of the full-length mRNA (type 14, P

    Article Snippet: Three probes were based on the common exon structure as found in full-length cDNA: exons 4 and 5 (type 9967; Hs00259967), 1 and 2 (type 10; Hs00938410_m1) and 5 and 6 (type 14; Hs00938414_m1); splice variant 5183, exons 2 and 5; and t-DARPP (type 16; Hs00938416_g1).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Amplification

    Genetic variation in PPP1R1B.

    Journal: Journal of Clinical Investigation

    Article Title: Genetic evidence implicating DARPP-32 in human frontostriatal structure, function, and cognition

    doi: 10.1172/JCI30413

    Figure Lengend Snippet: Genetic variation in PPP1R1B.

    Article Snippet: Three probes were based on the common exon structure as found in full-length cDNA: exons 4 and 5 (type 9967; Hs00259967), 1 and 2 (type 10; Hs00938410_m1) and 5 and 6 (type 14; Hs00938414_m1); splice variant 5183, exons 2 and 5; and t-DARPP (type 16; Hs00938416_g1).

    Techniques:

    Generation and characterization of Cre-expressing, HA-tagged human spinophilin mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Generation and characterization of Cre-expressing, HA-tagged human spinophilin mice. ( A ) A construct containing DNA encoding HA-tagged human spinophilin with a P2A sequence and mNeptune3 fluorescent protein was cloned into the pBIGT vector that contains a floxed-stop sequence. ( B ) The construct encoding the floxed-stop sequence and the HA-spinophilin-P2A-mNeptune 3 sequence was subcloned into the ROSA targeting vector pROSA_26.PA. ( C ) The modified ROSA vector was used for generation of the targeted transgenic mice. ( D ) Striatal cells were transfected without or with HA-tagged human spinophilin. Lysates were immunoprecipitated with either an HA or spinophilin antibody and immunoblotted with an HA antibody or a spinophilin antibody. HA-spinophilin was selectively detected when it was overexpressed. ( E ) Mice express HA-tagged spinophilin upon crossing with Cre recombinase expressed in the direct pathway (D1) or indirect pathway (A2A) medium spiny neurons. ( F ) Spinophilin and protein phosphatase 1 immunoblots of inputs and HA-immunoprecipitates from HA spinophilin mice crossed with D1 or A2A Cre-recombinase-expressing mice. ( G ) Mice expressing HA-spinophilin had non-significant increases in total spinophilin expression.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Expressing, Mouse Assay, Construct, Sequencing, Clone Assay, Plasmid Preparation, Modification, Transgenic Assay, Transfection, Immunoprecipitation, Western Blot

    Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Validation of spinophilin interactions. WT or spinophilin KO striatal (STR) or olfactory tubercle (OT) lysates were immunoprecipitated with a spinophilin antibody. Lysates or immunoprecipitates were immunoblotted for spinophilin and three interacting proteins that were detected in the HA immunoprecipitates that had a decreased (SAP102) or increased (Clathrin heavy chain and SRCIN1) interaction with spinophilin in amphetamine-treated animals. Spinophilin and all associated proteins were detected in the spinophilin immunoprecipitates from WT animals, but were absent in immunoprecipitates isolated from KO animals.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Immunoprecipitation, Isolation

    Greater abundance of spinophilin interacting proteins in amphetamine-treated animals occurs across both sexes and cell types. ( A ) Principal component analysis of individual samples normalized to spinophilin abundance within each sample and filtered for eight or more PSMs. ( B ) A volcano plot showing a majority of the proteins have increased abundance in HA immunoprecipitates isolated from amphetamine treated animals when normalized to the amphetamine-dependent increase in spinophilin abundance. ( C ) A plot of the abundance of spinophilin interacting proteins isolated from male, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. Left panel shows mean ± standard deviation, the right panel just shows the mean of the two values. ( D ) A plot of the abundance of spinophilin interacting proteins isolated from female, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. ( E ) A plot of the abundance of spinophilin interacting proteins isolated from female, A2A-Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples.

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Greater abundance of spinophilin interacting proteins in amphetamine-treated animals occurs across both sexes and cell types. ( A ) Principal component analysis of individual samples normalized to spinophilin abundance within each sample and filtered for eight or more PSMs. ( B ) A volcano plot showing a majority of the proteins have increased abundance in HA immunoprecipitates isolated from amphetamine treated animals when normalized to the amphetamine-dependent increase in spinophilin abundance. ( C ) A plot of the abundance of spinophilin interacting proteins isolated from male, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. Left panel shows mean ± standard deviation, the right panel just shows the mean of the two values. ( D ) A plot of the abundance of spinophilin interacting proteins isolated from female, D1 Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples. ( E ) A plot of the abundance of spinophilin interacting proteins isolated from female, A2A-Cre expressing animals and normalized for spinophilin expression ( Table S2 ) and quantified from treated (Y-axis) or control (X-axis) samples.

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Isolation, Expressing, Standard Deviation

    Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p

    Journal: Proteomes

    Article Title: Proteomic Analysis of the Spinophilin Interactome in Rodent Striatum Following Psychostimulant Sensitization

    doi: 10.3390/proteomes6040053

    Figure Lengend Snippet: Quantitation of spinophilin complexes isolated from dMSNs and iMSNs using tandem mass tag (TMT) analysis. ( A ) Striatal lysates isolated from male or female mice expressing HA spinophilin under the control of D1 or A2A promoters and treated with saline or amphetamine were immunoprecipitated with an HA antibody, digested with trypsin, labelled with eight different TMT tags, mixed and analyzed by mass spectrometry (MS/MS). ( B ) A higher intensity of TMT reporter abundance matching spinophilin was observed in amphetamine-treated compared to saline treated animals ( t -test; * p

    Article Snippet: Striatal cell lysates were immunoprecipitated with 1.6 µg of a sheep spinophilin antibody (ThermoFisher Scientific).

    Techniques: Quantitation Assay, Isolation, Mouse Assay, Expressing, Immunoprecipitation, Mass Spectrometry

    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 blot TIB73 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