phenylmethylsulfonyl fluouride  (Millipore)


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  • 99
    Name:
    Phenylmethanesulfonyl fluoride
    Description:

    Catalog Number:
    p7626
    Price:
    None
    Applications:
    Phenylmethanesulfonyl fluoride has been used in following applications:. cell fractionation.. used as a supplement in nuclear protein extraction.. inhibitor of cholesterol esterase (CE) and pseudocholinesterase (PCE).. used for the collection of blood prior to centrifugation to quantify plasma ANP levels.
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    Structured Review

    Millipore phenylmethylsulfonyl fluouride
    Phenylmethanesulfonyl fluoride

    https://www.bioz.com/result/phenylmethylsulfonyl fluouride/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phenylmethylsulfonyl fluouride - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: Intracellular Topology and Epitope Shielding of Poliovirus 3A Protein
    Article Snippet: .. Proteolysis was stopped by the addition of 20 μl of a solution containing 1 mM phenylmethylsulfonyl fluoride (Sigma) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer. ..

    Flow Cytometry:

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. Reagents HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    Protease Inhibitor:

    Article Title: Insights into the Structure of the Vip3Aa Insecticidal Protein by Protease Digestion Analysis
    Article Snippet: .. The irreversible inhibitors used to stop the trypsin action, were: PMSF (phenylmethanesulfonyl fluoride, from SIGMA), TLCK (Nα-tosyl-l -lysine chloromethyl ketone hydrochloride, from SIGMA), AEBSF protease inhibitor (from ThermoFisher, Waltham, MA, USA), and E64 (trans-epoxysuccinyl-l -leucylamido (4-guanidino) butane, from SIGMA). ..

    Purification:

    Article Title: The ORF45 Protein of Kaposi's Sarcoma-Associated Herpesvirus Is Associated with Purified Virions
    Article Snippet: .. Purified virions (10 μg of total protein) were treated with trypsin (4 μg/ml) (Promega, Madison, Wis.) in 100 μl of buffer (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 1 mM CaCl2 ) at 37°C for 1 h. Trypsin digestions were terminated by adding phenylmethylsulfonyl fluoride to a 0.5 mM concentration and a 1/100 volume of protease inhibitors (Sigma). .. In parallel experiments, Triton X-100 was added to a final of concentration of 1% to remove the viral envelope and expose the tegument nucleocapsid to the protease.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. Reagents HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    Concentration Assay:

    Article Title: The ORF45 Protein of Kaposi's Sarcoma-Associated Herpesvirus Is Associated with Purified Virions
    Article Snippet: .. Purified virions (10 μg of total protein) were treated with trypsin (4 μg/ml) (Promega, Madison, Wis.) in 100 μl of buffer (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 1 mM CaCl2 ) at 37°C for 1 h. Trypsin digestions were terminated by adding phenylmethylsulfonyl fluoride to a 0.5 mM concentration and a 1/100 volume of protease inhibitors (Sigma). .. In parallel experiments, Triton X-100 was added to a final of concentration of 1% to remove the viral envelope and expose the tegument nucleocapsid to the protease.

    other:

    Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction
    Article Snippet: 2.2 Materials Tyrphostin AG 1478, Phenylephrine, Apocynin, Peg-Catalase (PG-Cat), Dihydroethidium (DHE), phenanthroline, Phenylmethylsulfonyl fluoride were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Article Title: Matrix metalloproteinase-2-induced epidermal growth factor receptor transactivation impairs redox balance in vascular smooth muscle cells and facilitates vascular contraction
    Article Snippet: Tyrphostin AG 1478, Phenylephrine, Apocynin, Peg-Catalase (PG-Cat), Dihydroethidium (DHE), phenanthroline, Phenylmethylsulfonyl fluoride were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Article Title: Interrogating Parkinson's disease LRRK2 kinase pathway activity by assessing Rab10 phosphorylation in human neutrophils
    Article Snippet: The PF-06447475 inhibitor was purchased from R & D systems (#5716), diisopropylfluorophosphate (DIFP) was from Sigma (Cat# D0879) and phenylmethane sulfonyl fluoride (PMSF) was from Sigma (Cat# 78830).

    High Content Screening:

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    Article Title: Effects of hemocoagulase agkistrodon on the coagulation factors and its procoagulant activities
    Article Snippet: .. Reagents HCA (Beijing, Konruns Pharmaceutical Co., Ltd.); DEAE-Sepharose Fast Flow and Sephadex-G25 were obtained from Amersham Biosciences (Uppsala, Sweden); peroxidase-conjugated goat anti-rabbit IgG was purchased from BoaoBioTech (Beijing, People’s Republic of China); human fibrinogen, FPA, FPB, phenylmethanesulfonyl fluoride (PMSF), egg-yolk L-α-phosphatidylcholine, and bovine brain L-α-phosphatidylserine were from Sigma-Aldrich Co. (St Louis, MO, USA); human prothrombin (FII), thrombin, active factor V (FVa), FVII/VIIa, factor IX (FIX)/IXa, and FX/Xa were purchased from Hematologic Technologies (Burlington, VT, USA); the chromogenic substrate H-D-Phe-Pip-Arg-pNa•2HCl (S2238) was from Hyphen BioMed (Neuville-Sur-Oise, France); rabbit anti-HCA polyclonal antibodies were prepared in our laboratory as described in Supplementary materials (section “The preparation of polyclonal antibody”) and its ELISA titer was above 1:1,000,00. .. The specificity of polyclonal antibodies was confirmed by Western blot analysis (data not shown).

    SDS Page:

    Article Title: Intracellular Topology and Epitope Shielding of Poliovirus 3A Protein
    Article Snippet: .. Proteolysis was stopped by the addition of 20 μl of a solution containing 1 mM phenylmethylsulfonyl fluoride (Sigma) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer. ..

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  • 99
    Millipore pmsf
    Expression of a 120-kDa gelatinase by Id-1-expressing mammary epithelial cells. (A) Gelatin zymogram of conditioned media from SCp2–Id-1A (lane 1) and SCp2–Id-1E (lane 2) cells. Cells were cultured in serum-free medium, and conditioned media were harvested and analyzed on a gelatin substrate gel, as described in Materials and Methods. (B) Gelatin zymogram of conditioned media from SCp2–Id-1A cells either growth arrested by serum deprivation (lane 1) or growing in 5% serum (lane 2). (C) Gelatin zymogram of conditioned media from control SCp2 cells (lane 1) and an uncloned SCp2–Id-1-transfected pooled population (lane 2). Cells were cultured in serum-free medium. (D) Gelatin zymogram of SCp2–Id-1A (lanes 1, 3, and 5) and SCp2–Id-1E (lanes 2, 4, and 6) cell-conditioned media incubated with <t>DMSO</t> (lanes 1 and 2), the MMP inhibitor GM6001 (0.2 mM in DMSO) (lanes 3 and 4), or the serine proteinase inhibitor <t>PMSF</t> (5 mM in DMSO) (lanes 5 and 6). (E) Casein zymogram of conditioned media from SCp2–Id-1A (lanes 1 and 3) and SCp2–Id-1E (lanes 2 and 4) cells incubated with DMSO (lanes 1 and 2) or GM6001 (lanes 3 and 4). In panels A through D, arrows mark the positions of the 120-kDa MMP.
    Pmsf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3381 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmsf/product/Millipore
    Average 99 stars, based on 3381 article reviews
    Price from $9.99 to $1999.99
    pmsf - by Bioz Stars, 2020-07
    99/100 stars
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    99
    Millipore hat assay buffer
    Inhibitory effects of TA on <t>HAT</t> activity. (A) A colorimetric assay was performed to measure HAT activity in the presence of TA at the indicated concentrations. The results are presented as percentages relative to the control sample, which was incubated without TA (left panel). The IC 50 value of TA inhibition of HAT activity was calculated using Prism software (right panel). (B) An in vitro HAT assay using autoradiography was performed to evaluate the effect of the anti-HAT activity of TA on the acetylation of a synthetic H4 peptide by a <t>HeLa</t> NE. The arrow indicates histone H4 (left panel). The autoradiography was quantified using Image J software (right panel). The values presented are the means ± SD of two independent experiments. Means with different superscript letters are significantly different, p
    Hat Assay Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hat assay buffer/product/Millipore
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    hat assay buffer - by Bioz Stars, 2020-07
    99/100 stars
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    Expression of a 120-kDa gelatinase by Id-1-expressing mammary epithelial cells. (A) Gelatin zymogram of conditioned media from SCp2–Id-1A (lane 1) and SCp2–Id-1E (lane 2) cells. Cells were cultured in serum-free medium, and conditioned media were harvested and analyzed on a gelatin substrate gel, as described in Materials and Methods. (B) Gelatin zymogram of conditioned media from SCp2–Id-1A cells either growth arrested by serum deprivation (lane 1) or growing in 5% serum (lane 2). (C) Gelatin zymogram of conditioned media from control SCp2 cells (lane 1) and an uncloned SCp2–Id-1-transfected pooled population (lane 2). Cells were cultured in serum-free medium. (D) Gelatin zymogram of SCp2–Id-1A (lanes 1, 3, and 5) and SCp2–Id-1E (lanes 2, 4, and 6) cell-conditioned media incubated with DMSO (lanes 1 and 2), the MMP inhibitor GM6001 (0.2 mM in DMSO) (lanes 3 and 4), or the serine proteinase inhibitor PMSF (5 mM in DMSO) (lanes 5 and 6). (E) Casein zymogram of conditioned media from SCp2–Id-1A (lanes 1 and 3) and SCp2–Id-1E (lanes 2 and 4) cells incubated with DMSO (lanes 1 and 2) or GM6001 (lanes 3 and 4). In panels A through D, arrows mark the positions of the 120-kDa MMP.

    Journal: Molecular and Cellular Biology

    Article Title: A Novel Pathway for Mammary Epithelial Cell Invasion Induced by the Helix-Loop-Helix Protein Id-1

    doi:

    Figure Lengend Snippet: Expression of a 120-kDa gelatinase by Id-1-expressing mammary epithelial cells. (A) Gelatin zymogram of conditioned media from SCp2–Id-1A (lane 1) and SCp2–Id-1E (lane 2) cells. Cells were cultured in serum-free medium, and conditioned media were harvested and analyzed on a gelatin substrate gel, as described in Materials and Methods. (B) Gelatin zymogram of conditioned media from SCp2–Id-1A cells either growth arrested by serum deprivation (lane 1) or growing in 5% serum (lane 2). (C) Gelatin zymogram of conditioned media from control SCp2 cells (lane 1) and an uncloned SCp2–Id-1-transfected pooled population (lane 2). Cells were cultured in serum-free medium. (D) Gelatin zymogram of SCp2–Id-1A (lanes 1, 3, and 5) and SCp2–Id-1E (lanes 2, 4, and 6) cell-conditioned media incubated with DMSO (lanes 1 and 2), the MMP inhibitor GM6001 (0.2 mM in DMSO) (lanes 3 and 4), or the serine proteinase inhibitor PMSF (5 mM in DMSO) (lanes 5 and 6). (E) Casein zymogram of conditioned media from SCp2–Id-1A (lanes 1 and 3) and SCp2–Id-1E (lanes 2 and 4) cells incubated with DMSO (lanes 1 and 2) or GM6001 (lanes 3 and 4). In panels A through D, arrows mark the positions of the 120-kDa MMP.

    Article Snippet: Concentrated conditioned medium was mixed with nonreducing Laemmli sample buffer and incubated at 37°C for 15 min. After electrophoresis, the gels were incubated for 1 h in 2.5% Triton X-100 at room temperature, followed by 24 to 48 h in substrate buffer (100 mM Tris-HCl [pH 7.4]–15 mM CaCl2 ) in the absence or presence of GM6001 (0.2 mM in DMSO; supplied by Glycomed Corporation and obtained from Z. Werb [ ]), EDTA (10 mM), ortho -phenanthroline (1 mM in DMSO; Sigma), PMSF (5 mM), or AEBSF (0.5 mM; Calbiochem).

    Techniques: Expressing, Cell Culture, Transfection, Incubation

    Expression MosIR binding by dsRBPs TARBP and PACT. (A) Western blot showing expression and immunoprecipitation of HA-tagged TARBP2 and PACT proteins using anti-HA antibody. For TARBP2 and PACT immunoprecipitation, cultured cells were collected into 2 ml of PBS 48 h after transfection. The samples were centrifuged at maximal speed for 1 min, and the supernatants were removed. The cells were lysed with 200 μl of lysis buffer (20 mM Hepes [pH 7.8], 100 mM NaCl, 1 mM EDTA [pH 8.0], 0.5% IGEPAL-25%, 1 mM fresh DTT, 0.5 mM PMSF, 1 mM NaF, 0.2 mM Na 3 VO 4 , supplemented with 2× protease inhibitor cocktail set [Millipore], 2× phosphatase inhibitor cocktail set [Millipore], and RiboLock RNase inhibitor [Thermo Fisher Scientific]), passed through a needle and stored at ice for 10–15 min. The samples were centrifuged (15 min, 4°C, 12,000 g ), and the supernatants were used for immunoprecipitation. The supernatants were diluted with binding buffer (20 mM Hepes [pH 7.8], 100 mM NaCl, 1 mM EDTA [pH 8.0], 1 mM fresh DTT, 0.5 mM PMSF, 1 mM NaF, 0.2 mM Na 3 VO 4 , 2× protease inhibitor cocktail set [Millipore], 2× phosphatase inhibitor cocktail set [Millipore], and RiboLock RNase inhibitor [Thermo Fisher Scientific]) to 0.1% total concentration of IGEPAL-25. HA-probe antibody (F-7, Santa Cruz Biotech., sc-7392 AC) conjugated with agarose beads was used for the protein immunoprecipitation. The beads were added to each sample and incubated on rotator overnight at 4°C. The beads were washed with 1,000 μl of binding buffer 5 times (centrifugation at 4°C, 4,000 g , and 4 min). The last wash was placed into a new tube. Immunoprecipitated RNA was quantified using RT-qPCR. (B) Relative enrichment of CAG-EGFP-MosIR RNA upon immunoprecipitation of TARBP2 and PACT.

    Journal: Life Science Alliance

    Article Title: Main constraints for RNAi induced by expressed long dsRNA in mouse cells

    doi: 10.26508/lsa.201800289

    Figure Lengend Snippet: Expression MosIR binding by dsRBPs TARBP and PACT. (A) Western blot showing expression and immunoprecipitation of HA-tagged TARBP2 and PACT proteins using anti-HA antibody. For TARBP2 and PACT immunoprecipitation, cultured cells were collected into 2 ml of PBS 48 h after transfection. The samples were centrifuged at maximal speed for 1 min, and the supernatants were removed. The cells were lysed with 200 μl of lysis buffer (20 mM Hepes [pH 7.8], 100 mM NaCl, 1 mM EDTA [pH 8.0], 0.5% IGEPAL-25%, 1 mM fresh DTT, 0.5 mM PMSF, 1 mM NaF, 0.2 mM Na 3 VO 4 , supplemented with 2× protease inhibitor cocktail set [Millipore], 2× phosphatase inhibitor cocktail set [Millipore], and RiboLock RNase inhibitor [Thermo Fisher Scientific]), passed through a needle and stored at ice for 10–15 min. The samples were centrifuged (15 min, 4°C, 12,000 g ), and the supernatants were used for immunoprecipitation. The supernatants were diluted with binding buffer (20 mM Hepes [pH 7.8], 100 mM NaCl, 1 mM EDTA [pH 8.0], 1 mM fresh DTT, 0.5 mM PMSF, 1 mM NaF, 0.2 mM Na 3 VO 4 , 2× protease inhibitor cocktail set [Millipore], 2× phosphatase inhibitor cocktail set [Millipore], and RiboLock RNase inhibitor [Thermo Fisher Scientific]) to 0.1% total concentration of IGEPAL-25. HA-probe antibody (F-7, Santa Cruz Biotech., sc-7392 AC) conjugated with agarose beads was used for the protein immunoprecipitation. The beads were added to each sample and incubated on rotator overnight at 4°C. The beads were washed with 1,000 μl of binding buffer 5 times (centrifugation at 4°C, 4,000 g , and 4 min). The last wash was placed into a new tube. Immunoprecipitated RNA was quantified using RT-qPCR. (B) Relative enrichment of CAG-EGFP-MosIR RNA upon immunoprecipitation of TARBP2 and PACT.

    Article Snippet: Before collection, the cells were washed with PBS and lysed in lysis buffer (20 mM Hepes [pH 7.8], 100 mM NaCl, 1 mM EDTA [pH 8.0], 0.5% IGEPAL-25%, 1 mM fresh DTT, 0.5 mM PMSF, 1 mM NaF, 0.2 mM Na3 VO4 , supplemented with 2× protease inhibitor cocktail set [Millipore], 2× phosphatase inhibitor cocktail set [Millipore], and RiboLock RNase inhibitor [Thermo Fisher Scientific]).

    Techniques: Expressing, Binding Assay, Western Blot, Immunoprecipitation, Cell Culture, Transfection, Lysis, Protease Inhibitor, Concentration Assay, Incubation, Centrifugation, Quantitative RT-PCR

    Inhibitory effects of TA on HAT activity. (A) A colorimetric assay was performed to measure HAT activity in the presence of TA at the indicated concentrations. The results are presented as percentages relative to the control sample, which was incubated without TA (left panel). The IC 50 value of TA inhibition of HAT activity was calculated using Prism software (right panel). (B) An in vitro HAT assay using autoradiography was performed to evaluate the effect of the anti-HAT activity of TA on the acetylation of a synthetic H4 peptide by a HeLa NE. The arrow indicates histone H4 (left panel). The autoradiography was quantified using Image J software (right panel). The values presented are the means ± SD of two independent experiments. Means with different superscript letters are significantly different, p

    Journal: Molecular Metabolism

    Article Title: Tannic acid, a novel histone acetyltransferase inhibitor, prevents non-alcoholic fatty liver disease both in vivo and in vitro model

    doi: 10.1016/j.molmet.2018.11.001

    Figure Lengend Snippet: Inhibitory effects of TA on HAT activity. (A) A colorimetric assay was performed to measure HAT activity in the presence of TA at the indicated concentrations. The results are presented as percentages relative to the control sample, which was incubated without TA (left panel). The IC 50 value of TA inhibition of HAT activity was calculated using Prism software (right panel). (B) An in vitro HAT assay using autoradiography was performed to evaluate the effect of the anti-HAT activity of TA on the acetylation of a synthetic H4 peptide by a HeLa NE. The arrow indicates histone H4 (left panel). The autoradiography was quantified using Image J software (right panel). The values presented are the means ± SD of two independent experiments. Means with different superscript letters are significantly different, p

    Article Snippet: For autoradiography-based in vitro HAT activity assays, HeLa cell nuclear extract was incubated with HAT assay buffer (50 mM HEPES, pH 8.0; 10% glycerol; 1 mM DTT; 1 mM phenylmethylsulfonyl fluoride; and 10 mM sodium butyrate), 1 μL [3 H] acetyl-CoA, and 5 μg of biotinylated-H4 peptide (Millipore, Billerica, MA, USA) along with TA at the indicated concentrations at 30 °C for 1 h. The reactions were stopped by adding 5 × sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE) sample buffer and samples were separated on 15% SDS–PAGE gels, followed by autoradiographic analysis.

    Techniques: HAT Assay, Activity Assay, Colorimetric Assay, Incubation, Inhibition, Software, In Vitro, Autoradiography

    Inhibitory effects of TA on HAT activity. (A) A colorimetric assay was performed to measure HAT activity in the presence of TA at the indicated concentrations. The results are presented as percentages relative to the control sample, which was incubated without TA (left panel). The IC 50 value of TA inhibition of HAT activity was calculated using Prism software (right panel). (B) An in vitro HAT assay using autoradiography was performed to evaluate the effect of the anti-HAT activity of TA on the acetylation of a synthetic H4 peptide by a HeLa NE. The arrow indicates histone H4 (left panel). The autoradiography was quantified using Image J software (right panel). The values presented are the means ± SD of two independent experiments. Means with different superscript letters are significantly different, p

    Journal: Molecular Metabolism

    Article Title: Tannic acid, a novel histone acetyltransferase inhibitor, prevents non-alcoholic fatty liver disease both in vivo and in vitro model

    doi: 10.1016/j.molmet.2018.11.001

    Figure Lengend Snippet: Inhibitory effects of TA on HAT activity. (A) A colorimetric assay was performed to measure HAT activity in the presence of TA at the indicated concentrations. The results are presented as percentages relative to the control sample, which was incubated without TA (left panel). The IC 50 value of TA inhibition of HAT activity was calculated using Prism software (right panel). (B) An in vitro HAT assay using autoradiography was performed to evaluate the effect of the anti-HAT activity of TA on the acetylation of a synthetic H4 peptide by a HeLa NE. The arrow indicates histone H4 (left panel). The autoradiography was quantified using Image J software (right panel). The values presented are the means ± SD of two independent experiments. Means with different superscript letters are significantly different, p

    Article Snippet: For autoradiography-based in vitro HAT activity assays, HeLa cell nuclear extract was incubated with HAT assay buffer (50 mM HEPES, pH 8.0; 10% glycerol; 1 mM DTT; 1 mM phenylmethylsulfonyl fluoride; and 10 mM sodium butyrate), 1 μL [3 H] acetyl-CoA, and 5 μg of biotinylated-H4 peptide (Millipore, Billerica, MA, USA) along with TA at the indicated concentrations at 30 °C for 1 h. The reactions were stopped by adding 5 × sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE) sample buffer and samples were separated on 15% SDS–PAGE gels, followed by autoradiographic analysis.

    Techniques: HAT Assay, Activity Assay, Colorimetric Assay, Incubation, Inhibition, Software, In Vitro, Autoradiography