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, 2021-06
    99/100 stars

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    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.

    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: Altered Levels of Toll-Like Receptors in Circulating Extracellular Vesicles in Multiple Sclerosis
    Article Snippet: For the NTA, the particle concentration in function of particle diameter was averaged from five 20-s videos captured using the following settings: scattering camera level, 15; slider shutter, 1206; slider gain, 366; detection threshold, 3; and sample dilution ranging from 1:100 to 1:200 providing a range of 50–250 particles/frame. .. The remaining volume was mixed with 300 μL of MPER lysis buffer (Thermo Scientific, Grand Island, NY, U.S.) supplemented with protease (cOmplete ULTRA Tablets, Roche, Millipore Sigma, St. Louis, MO, U.S.) and phosphatase (HALT Phosphatase Inhibitor Cocktail, Thermo Fisher Scientific, Cincinnati, OH, U.S.) inhibitors and was subjected to two freeze-thaw cycles to generate lysates for downstream ELISA. .. EV Array The EV Array was developed based on the technology of protein microarray to phenotype serum-derived EVs (optimized for small EVs/exosomes) for multiple antigens without any enrichment or purification prior to analysis [ ].

    Article Title: Ultrasound with microbubbles improves memory, ameliorates pathology and modulates hippocampal proteomic changes in a triple transgenic mouse model of Alzheimer's disease
    Article Snippet: Western blot analysisTo confirm the differentially expressed proteins revealed by 2D-DIGE, the expression of the proteins was further measured by western-blot analysis. .. Hippocampal proteins from WT mice, as well as 3×Tg-AD mice that received either sham or FUS/MB treatment, were extracted using lysis buffer (Beyotime, China) with protease and phosphatase inhibitor cocktail (Thermo Scientific, USA). .. Then the supernatant was collected and the protein concentration was estimated using micro BCA protein assay kit (Thermo Scientific, USA).

    Article Title: Increased expression of the mitochondrial derived peptide, MOTS-c, in skeletal muscle of healthy aging men is associated with myofiber composition
    Article Snippet: .. Immunoblotting ~20 mg of muscle was homogenized for 40s at 20 Hz using a TissueLyser (Qiagen, Venlo, Netherlands) in lysis buffer containing 10 μL/mg 25 mM Tris 0.5% v/v Triton X-100 supplemented with Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Scientific, Waltham, MA). .. The supernatant was collected after centrifugation to determine protein concentration using the BCA protein assay kit (Thermo Scientific, Waltham, MA).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Altered Levels of Toll-Like Receptors in Circulating Extracellular Vesicles in Multiple Sclerosis
    Article Snippet: For the NTA, the particle concentration in function of particle diameter was averaged from five 20-s videos captured using the following settings: scattering camera level, 15; slider shutter, 1206; slider gain, 366; detection threshold, 3; and sample dilution ranging from 1:100 to 1:200 providing a range of 50–250 particles/frame. .. The remaining volume was mixed with 300 μL of MPER lysis buffer (Thermo Scientific, Grand Island, NY, U.S.) supplemented with protease (cOmplete ULTRA Tablets, Roche, Millipore Sigma, St. Louis, MO, U.S.) and phosphatase (HALT Phosphatase Inhibitor Cocktail, Thermo Fisher Scientific, Cincinnati, OH, U.S.) inhibitors and was subjected to two freeze-thaw cycles to generate lysates for downstream ELISA. .. EV Array The EV Array was developed based on the technology of protein microarray to phenotype serum-derived EVs (optimized for small EVs/exosomes) for multiple antigens without any enrichment or purification prior to analysis [ ].

    Protease Inhibitor:

    Article Title: Antigen Presentation Profiling Reveals Recognition of Lymphoma Immunoglobulin Neoantigens
    Article Snippet: Total Cell Line Proteome Sample Preparation and high pH Reversed Phase Fractionation for MS-analysis 1x108 cells from each of two MCL cell lines (JEKO and L128) were used for protein extractions. .. In brief, cells were lysed in 8M urea, 150 mM NaCl, 5 mM DTT, 50 mM Tris pH 8 supplemented with Complete Protease Inhibitor Cocktail tablet (Roche) and 1x Halt™ Protease and Phosphatase Inhibitor Cocktail (ThermoFisher Scientific). .. Following lysis, the lysate was centrifuged at 13,200 rpm for 15 min, the supernatant was transferred to fresh tubes and a second round of centrifugation was performed.

    Mouse Assay:

    Article Title: Ultrasound with microbubbles improves memory, ameliorates pathology and modulates hippocampal proteomic changes in a triple transgenic mouse model of Alzheimer's disease
    Article Snippet: Western blot analysisTo confirm the differentially expressed proteins revealed by 2D-DIGE, the expression of the proteins was further measured by western-blot analysis. .. Hippocampal proteins from WT mice, as well as 3×Tg-AD mice that received either sham or FUS/MB treatment, were extracted using lysis buffer (Beyotime, China) with protease and phosphatase inhibitor cocktail (Thermo Scientific, USA). .. Then the supernatant was collected and the protein concentration was estimated using micro BCA protein assay kit (Thermo Scientific, USA).

    Modification:

    Article Title: Membrane-initiated estradiol signaling in immortalized hypothalamic N-38 neurons
    Article Snippet: .. Cells were washed with ice cold TBS and collected in a modified RIPA buffer (Triton X-100 1.25%, SDS 0.1%, 50 mM Tris-HCl, 150 mM NaCl, 5 mM EDTA, pH 7.6) supplemented with Halt™ protease and phosphatase inhibitor cocktails (Thermo). .. An aliquot of the supernatant was taken for protein determination using the BCA method (Thermo) and for examination of cytoplasmic proteins PKCθ and β-actin (the latter was used for western blotting loading control).

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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, 2021-06
    97/100 stars
      Buy from Supplier

    Image Search Results


    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