bovine serum albumin bsa  (Millipore)


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    Structured Review

    Millipore bovine serum albumin bsa
    12R‐ LOX activity was blocked in PE cheek and PP post‐auricular samples of the second and eighth tape strippings with a <t>polyclonal</t> 12R‐ LOX antibody. About 0.5% <t>BSA</t> was used as the control. Data are shown as mean ± SD , n = 6 * P ≤ 0.05, ** P ≤ 0.01, or *** P ≤ 0.0001.
    Bovine Serum Albumin Bsa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2302 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation"

    Article Title: 12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation

    Journal: International Journal of Cosmetic Science

    doi: 10.1111/ics.12532

    12R‐ LOX activity was blocked in PE cheek and PP post‐auricular samples of the second and eighth tape strippings with a polyclonal 12R‐ LOX antibody. About 0.5% BSA was used as the control. Data are shown as mean ± SD , n = 6 * P ≤ 0.05, ** P ≤ 0.01, or *** P ≤ 0.0001.
    Figure Legend Snippet: 12R‐ LOX activity was blocked in PE cheek and PP post‐auricular samples of the second and eighth tape strippings with a polyclonal 12R‐ LOX antibody. About 0.5% BSA was used as the control. Data are shown as mean ± SD , n = 6 * P ≤ 0.05, ** P ≤ 0.01, or *** P ≤ 0.0001.

    Techniques Used: Activity Assay

    12R‐ LOX activity was not blocked by the polyclonal eLOX 3 antibody. 0.5% BSA was used the control. Data are shown as mean ± SD , n = 6. No significant differences were found.
    Figure Legend Snippet: 12R‐ LOX activity was not blocked by the polyclonal eLOX 3 antibody. 0.5% BSA was used the control. Data are shown as mean ± SD , n = 6. No significant differences were found.

    Techniques Used: Activity Assay

    Related Articles

    Luciferase:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). .. CR9506 (3.66 mg/mL), CR9501-biotin (9.91 mg/mL), CR9506-biotin (3.10 mg/mL), and RSV CL57V224 labeled with firefly luciferase (RSV CL57V224-FFL) were obtained from Janssen Infectious Diseases and Vaccines (Leiden, The Netherlands).

    Article Title: Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses
    Article Snippet: Huh7-Lunet cells were washed with PBS and then incubated with a buffer consisting of 20 mM Tris-HCl pH 6.8, 50 mM NaCl, 4 mM CaCl2 , 0.01% bovine serum albumin (BSA) (mock treated) or the same buffer containing 0.5 U/ml heparinase I (Sigma-Aldrich), or heparinase III (Sigma-Aldrich) or chondroitinase ABC (Sigma-Aldrich). .. After incubation for 1 h at 37°C, cells were washed three times with PBS and then inoculated with 500 μl of reporter virus preparation for 1 h. Forty-eight hours later, cells were lysed in 350 μl of lysis buffer, and luciferase activity was measured as described above.

    Bradford Protein Assay:

    Article Title: In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung
    Article Snippet: Bradford protein assay reagent and PVDF Sequi-blot membranes were obtained from Bio-Rad (Hercules, CA). .. Bovine serum albumin (BSA), thiourea, tributylphosphine (TBP), and dithiothreitol (DTT) were ordered from Sigma-Aldrich (St. Louis, MO).

    Synthesized:

    Article Title: In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung
    Article Snippet: Bovine serum albumin (BSA), thiourea, tributylphosphine (TBP), and dithiothreitol (DTT) were ordered from Sigma-Aldrich (St. Louis, MO). .. IsoGel low EEO agarose was ordered from BMA (Rockland, ME), 14 C-1-NN and 1-NN were synthesized as previously described ( ).

    Immunostaining:

    Article Title: 12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation
    Article Snippet: .. The anti‐12R‐LOX and eLOX3 polyclonal antibodies used in the immunostaining were diluted in purged 0.5% bovine serum albumin (BSA) in reaction buffer as well as 5‐(methylamino)‐2‐(1‐naphthalenyl)‐4‐oxazolecarbonitrile (ML351) (Sigma‐Aldrich) as a blocker for the LOX enzymes. .. 12R‐LOX activity was expressed as U min−1 normalized to the total SC protein on tape strips.

    Electrophoresis:

    Article Title: Vesicular Glutamate and GABA Transporters Sort to Distinct Sets of Vesicles in a Population of Presynaptic Terminals
    Article Snippet: 10% polyacrylamide Criterion precast gels, nitrocellulose sheets (ECL), electrophoresis equipment and molecular mass markers (Precision plus) were from Biorad (Hercules, CA). .. Components of Durcupan ACM, sodium azide (NaN3 ), diaminobenzidine (DAB), human serum albumin (HSA), bovine serum albumin (BSA), sodium ethylenediamine tetraacetate (EDTA), polyethylene glycol (PEG), Trizma base and Tris–HCl were obtained from Sigma Aldrich (Oslo, Norway).

    Article Title: In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung
    Article Snippet: Bovine serum albumin (BSA), thiourea, tributylphosphine (TBP), and dithiothreitol (DTT) were ordered from Sigma-Aldrich (St. Louis, MO). .. All other electrophoresis materials and equipment were obtained from Amersham Biosciences (Piscataway, NJ).

    Ex Vivo:

    Article Title: Separation of the convulsions and antidepressant-like effects produced by the delta-opioid agonist SNC80 in rats
    Article Snippet: Paragraph title: Ex vivo [3 H]NTI-binding experiments ... Known concentrations of SNC80 diluted in 0.1% bovine serum albumin (BSA) were added to the assay tubes with Sigmacote-coated (Sigma, St. Louis, MO) pipette tips.

    Incubation:

    Article Title: Primary cilia mediate mitochondrial stress responses to promote dopamine neuron survival in a Parkinson’s disease model
    Article Snippet: .. Subsequently, the cells were blocked with PBS containing 1% bovine serum albumin (BSA), and incubated overnight at 4 °C with primary antibodies against acetylated α-tubulin (1:1000, T7451, Sigma-Aldrich) or ARL13B (1:1000, 17711-1-AP, Proteintech) in 1% BSA. .. After washing, the cells were incubated with Alexa Fluor 488 or 555-conjugated secondary antibodies at room temperature (RT) for 1 h. Before mounting, the cells were treated with Hoechst 33342 dye (1:10,000, H3570, Thermo-Fisher) for nuclear staining.

    Article Title: Evidence for the multimeric structure of ferroportin
    Article Snippet: .. Immunofluorescence was performed as previously described, with the following changes: after formaldehyde fixations, cells were incubated in phosphate-buffered saline (PBS), and 1% bovine serum albumin (BSA) with or without 0.1% saponin (Sigma, St Louis, MO). .. In the absence of detergent/permeabilization, only epitopes that are extracellular will be detected by immunofluorescence.

    Article Title: 12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation
    Article Snippet: Each tape was incubated with 0.1% Tergitol™‐NP40 in 50 mmol L−1 Tris (Sigma‐Aldrich) with Pierce™ Protease Inhibitor Tablets (1 per 10 mL; Thermo Fisher Scientific) for 20 min on a shaker with 600 rpm at 4°C. .. The anti‐12R‐LOX and eLOX3 polyclonal antibodies used in the immunostaining were diluted in purged 0.5% bovine serum albumin (BSA) in reaction buffer as well as 5‐(methylamino)‐2‐(1‐naphthalenyl)‐4‐oxazolecarbonitrile (ML351) (Sigma‐Aldrich) as a blocker for the LOX enzymes.

    Article Title: Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses
    Article Snippet: .. Huh7-Lunet cells were washed with PBS and then incubated with a buffer consisting of 20 mM Tris-HCl pH 6.8, 50 mM NaCl, 4 mM CaCl2 , 0.01% bovine serum albumin (BSA) (mock treated) or the same buffer containing 0.5 U/ml heparinase I (Sigma-Aldrich), or heparinase III (Sigma-Aldrich) or chondroitinase ABC (Sigma-Aldrich). .. After incubation for 1 h at 37°C, cells were washed three times with PBS and then inoculated with 500 μl of reporter virus preparation for 1 h. Forty-eight hours later, cells were lysed in 350 μl of lysis buffer, and luciferase activity was measured as described above.

    Activity Assay:

    Article Title: 12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation
    Article Snippet: The anti‐12R‐LOX and eLOX3 polyclonal antibodies used in the immunostaining were diluted in purged 0.5% bovine serum albumin (BSA) in reaction buffer as well as 5‐(methylamino)‐2‐(1‐naphthalenyl)‐4‐oxazolecarbonitrile (ML351) (Sigma‐Aldrich) as a blocker for the LOX enzymes. .. 12R‐LOX activity was expressed as U min−1 normalized to the total SC protein on tape strips.

    Article Title: Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses
    Article Snippet: Huh7-Lunet cells were washed with PBS and then incubated with a buffer consisting of 20 mM Tris-HCl pH 6.8, 50 mM NaCl, 4 mM CaCl2 , 0.01% bovine serum albumin (BSA) (mock treated) or the same buffer containing 0.5 U/ml heparinase I (Sigma-Aldrich), or heparinase III (Sigma-Aldrich) or chondroitinase ABC (Sigma-Aldrich). .. After incubation for 1 h at 37°C, cells were washed three times with PBS and then inoculated with 500 μl of reporter virus preparation for 1 h. Forty-eight hours later, cells were lysed in 350 μl of lysis buffer, and luciferase activity was measured as described above.

    Giemsa Stain:

    Article Title: DNA damage and homologous recombination signaling induced by thymidylate deprivation
    Article Snippet: .. Hydroxyurea (HU), Bovine albumin (BSA), thiazolyl blue tetrazolium bromide (MTT), β-glycerophosphate, β-mercaptoethanol (BME), phenylmethanesulfonyl fluoride (PMSF), sodium fluoride, sodium bicarbonate, dimethyl sulfoxide (DMSO), Giemsa Stain, and sodium othovanadate were purchased from Sigma (St. Louis, MO). .. Raltitrexed (RTX) was generously provided by AstraZeneca (U.K.).

    Expressing:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents. .. The relative levels of each protein expression were determined by densitometric analysis by using ImageJ software.

    BIA-KA:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: The protein concentrations were determined by a BCA kit (Pierce, Rockford, USA). .. After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents.

    Modification:

    Article Title: Involvement of the KIT/KITL Signaling Pathway in 4-Vinylcyclohexene Diepoxide-Induced Ovarian Follicle Loss in Rats 1
    Article Snippet: VCD (mixture of isomers, purity > 99%), collagenase (clostridium histolytic type I), DNase I, bovine serum albumin (BSA), Sigmacote, ascorbic acid (vitamin C), transferrin, d-biotin, Ribonuclease A, and all other unspecified chemicals were purchased from Sigma-Aldrich Inc. (St. Louis, MO). .. Dulbecco modified Eagle medium/nutrient mixture F-12 (Ham) 1× (DMEM/Ham F-12), medium 199 (M199), Albumax, penicillin/streptomycin (5000 U/ml, 5000 μg/ml), Hanks Balanced Salt Solution (without CaCl2 , MgCl2 , or MgSO4 ), Superscript III reverse transcriptase, oligo(dT)20 primers, MgCl2 (25 mM), Cot-1 DNA, and PBS were purchased from Invitrogen (Carlsbad, CA).

    Article Title: Preservation and Expansion of the Primate Keratocyte Phenotype by Downregulating TGF-? Signaling in a Low-Calcium, Serum-Free Medium
    Article Snippet: The tissue culture plastic plates (96-well and 6-well) were purchased from BD Biosciences (Lincoln Park, NJ); collagenase A and dispase II powder from Roche (Indianapolis, IN); amphotericin B, Dulbecco’s modified Eagle’s medium (DMEM), defined keratinocyte serum-free-medium (KSFM), FBS, gentamicin, Hanks ‘ balanced salt solution (HBSS), HEPES buffer, phosphate-buffered saline (PBS), soybean trypsin inhibitor, and 0.25% trypsin/1 mM EDTA were purchased from Invitrogen-Gibco (Grand Island, NY); A cell-viability–cytotoxicity kit (Live/Dead) from Invitrogen (Eugene, OR); endo-β-galactosidase was from Seikagaku (Tokyo, Japan); optimal cutting temperature (OCT) from Sakura Finetek (Torrance, CA); and a cell-proliferation assay (MTT) from Roche. .. Other reagents and chemicals including bovine serum albumin (BSA) and insulin-transferrin-sodium selenite (ITS) medium supplement were from Sigma-Aldrich.

    Western Blot:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: Paragraph title: Western blotting ... After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents.

    Electron Microscopy:

    Article Title: Vesicular Glutamate and GABA Transporters Sort to Distinct Sets of Vesicles in a Population of Presynaptic Terminals
    Article Snippet: Fluoromount G (water base), uranyl acetate, lead citrate, glutaraldehyde (GA), and nickel grids (for electron microscopy [EM]) were from Electron Microscopy Science (Washington, PA). .. Components of Durcupan ACM, sodium azide (NaN3 ), diaminobenzidine (DAB), human serum albumin (HSA), bovine serum albumin (BSA), sodium ethylenediamine tetraacetate (EDTA), polyethylene glycol (PEG), Trizma base and Tris–HCl were obtained from Sigma Aldrich (Oslo, Norway).

    Protease Inhibitor:

    Article Title: In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung
    Article Snippet: CHAPS, protease inhibitor cocktail III, and Triton X-100 were acquired from Calbiochem (LaJolla, CA). .. Bovine serum albumin (BSA), thiourea, tributylphosphine (TBP), and dithiothreitol (DTT) were ordered from Sigma-Aldrich (St. Louis, MO).

    Article Title: 12R‐lipoxygenase activity is reduced in photodamaged facial stratum corneum. A novel activity assay indicates a key function in corneocyte maturation
    Article Snippet: Each tape was incubated with 0.1% Tergitol™‐NP40 in 50 mmol L−1 Tris (Sigma‐Aldrich) with Pierce™ Protease Inhibitor Tablets (1 per 10 mL; Thermo Fisher Scientific) for 20 min on a shaker with 600 rpm at 4°C. .. The anti‐12R‐LOX and eLOX3 polyclonal antibodies used in the immunostaining were diluted in purged 0.5% bovine serum albumin (BSA) in reaction buffer as well as 5‐(methylamino)‐2‐(1‐naphthalenyl)‐4‐oxazolecarbonitrile (ML351) (Sigma‐Aldrich) as a blocker for the LOX enzymes.

    Transferring:

    Article Title: Separation of the convulsions and antidepressant-like effects produced by the delta-opioid agonist SNC80 in rats
    Article Snippet: .. Known concentrations of SNC80 diluted in 0.1% bovine serum albumin (BSA) were added to the assay tubes with Sigmacote-coated (Sigma, St. Louis, MO) pipette tips. ..

    Recombinant:

    Article Title: Involvement of the KIT/KITL Signaling Pathway in 4-Vinylcyclohexene Diepoxide-Induced Ovarian Follicle Loss in Rats 1
    Article Snippet: VCD (mixture of isomers, purity > 99%), collagenase (clostridium histolytic type I), DNase I, bovine serum albumin (BSA), Sigmacote, ascorbic acid (vitamin C), transferrin, d-biotin, Ribonuclease A, and all other unspecified chemicals were purchased from Sigma-Aldrich Inc. (St. Louis, MO). .. Recombinant mouse KITL and human BMP4 were obtained from R & D Systems Inc. (Minneapolis, MN).

    Article Title: Initial synthesis and characterization of an immobilized heat shock protein 90 column for online determination of binding affinities
    Article Snippet: Recombinant human HSP90α ( ∼ 90% pure) was purchased from Stressgen Bioreagents (Ann Arbor, MI, U.S.A.). .. Bovine serum albumin (BSA), ammonium acetate, ATP, 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC), glutaraldehyde, glutaric acid, glycine, pyridine (99.8%), sodium azide, and Tris buffer were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, U.S.A.).

    Immunofluorescence:

    Article Title: Evidence for the multimeric structure of ferroportin
    Article Snippet: .. Immunofluorescence was performed as previously described, with the following changes: after formaldehyde fixations, cells were incubated in phosphate-buffered saline (PBS), and 1% bovine serum albumin (BSA) with or without 0.1% saponin (Sigma, St Louis, MO). .. In the absence of detergent/permeabilization, only epitopes that are extracellular will be detected by immunofluorescence.

    Molecular Weight:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: The purity of the protein was > 96% and the molecular weight of the protein was approximately 175 kDa (confirmed with non-reducing SDS-PAGE and size-exclusion chromatography). .. Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    MTT Assay:

    Article Title: DNA damage and homologous recombination signaling induced by thymidylate deprivation
    Article Snippet: .. Hydroxyurea (HU), Bovine albumin (BSA), thiazolyl blue tetrazolium bromide (MTT), β-glycerophosphate, β-mercaptoethanol (BME), phenylmethanesulfonyl fluoride (PMSF), sodium fluoride, sodium bicarbonate, dimethyl sulfoxide (DMSO), Giemsa Stain, and sodium othovanadate were purchased from Sigma (St. Louis, MO). .. Raltitrexed (RTX) was generously provided by AstraZeneca (U.K.).

    Article Title: Preservation and Expansion of the Primate Keratocyte Phenotype by Downregulating TGF-? Signaling in a Low-Calcium, Serum-Free Medium
    Article Snippet: The tissue culture plastic plates (96-well and 6-well) were purchased from BD Biosciences (Lincoln Park, NJ); collagenase A and dispase II powder from Roche (Indianapolis, IN); amphotericin B, Dulbecco’s modified Eagle’s medium (DMEM), defined keratinocyte serum-free-medium (KSFM), FBS, gentamicin, Hanks ‘ balanced salt solution (HBSS), HEPES buffer, phosphate-buffered saline (PBS), soybean trypsin inhibitor, and 0.25% trypsin/1 mM EDTA were purchased from Invitrogen-Gibco (Grand Island, NY); A cell-viability–cytotoxicity kit (Live/Dead) from Invitrogen (Eugene, OR); endo-β-galactosidase was from Seikagaku (Tokyo, Japan); optimal cutting temperature (OCT) from Sakura Finetek (Torrance, CA); and a cell-proliferation assay (MTT) from Roche. .. Other reagents and chemicals including bovine serum albumin (BSA) and insulin-transferrin-sodium selenite (ITS) medium supplement were from Sigma-Aldrich.

    Fluorescence:

    Article Title: Primary cilia mediate mitochondrial stress responses to promote dopamine neuron survival in a Parkinson’s disease model
    Article Snippet: Subsequently, the cells were blocked with PBS containing 1% bovine serum albumin (BSA), and incubated overnight at 4 °C with primary antibodies against acetylated α-tubulin (1:1000, T7451, Sigma-Aldrich) or ARL13B (1:1000, 17711-1-AP, Proteintech) in 1% BSA. .. Cilia images were observed using a fluorescence microscope.

    Magnetic Beads:

    Article Title: Vesicular Glutamate and GABA Transporters Sort to Distinct Sets of Vesicles in a Population of Presynaptic Terminals
    Article Snippet: Magnetic beads coated with protein A were from Dynal (Invitrogen) (Carlsbad, CA). .. Components of Durcupan ACM, sodium azide (NaN3 ), diaminobenzidine (DAB), human serum albumin (HSA), bovine serum albumin (BSA), sodium ethylenediamine tetraacetate (EDTA), polyethylene glycol (PEG), Trizma base and Tris–HCl were obtained from Sigma Aldrich (Oslo, Norway).

    Size-exclusion Chromatography:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: The purity of the protein was > 96% and the molecular weight of the protein was approximately 175 kDa (confirmed with non-reducing SDS-PAGE and size-exclusion chromatography). .. Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    Article Title: Co-Immobilization of Tri-Enzymes for the Conversion of Hydroxymethylfurfural to 2,5-Diformylfuran
    Article Snippet: .. Materials Galactose oxidase from Dactylium dendroides (500–1500 U/mg protein, one unit generates a ΔA425 of 1.0 per minute at pH 6.0 at 25 °C in a peroxidase and o-tolidine system), catalase from bovine liver (2000–5000 U/mg protein, one unit decomposes to 1.0 micromole of hydrogen peroxide per minute at pH 7.0 at 25 °C), horseradish peroxidase ( > 200 U/mg protein, one unit offers 1.0 mg purpurogallin from pyrogallolin after 20 sec at pH 6.0 at 20 °C), HMF, KBr, bovine serum albumin (BSA), choline chloride, and glycerol were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). .. 3-Methoxybenzyl alcohol, hydrogen peroxide solution (30 wt % in H2 O), pyrogallol, hydroxymethylfurfural (HMF), 2,5-diformylfuran (DFF), and SIGMAFAST DAB with Metal Enhancer Tablet Sets (DAB) were also supplied by Sigma-Aldrich Chemical Co. All chemicals and reagents were of analytical grade.

    Labeling:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). .. CR9506 (3.66 mg/mL), CR9501-biotin (9.91 mg/mL), CR9506-biotin (3.10 mg/mL), and RSV CL57V224 labeled with firefly luciferase (RSV CL57V224-FFL) were obtained from Janssen Infectious Diseases and Vaccines (Leiden, The Netherlands).

    Mouse Assay:

    Article Title: Dose-Related Differences in the Regional Pattern of Cannabinoid Receptor Adaptation and in Vivo Tolerance Development to ?9-Tetrahydrocannabinol
    Article Snippet: ICR mice (male, 24–30 g) were obtained from Harlan (Indianapolis, IN). .. Bovine serum albumin (BSA), guanosine 5′-3- O -(thio)triphosphate, GDP, and WIN55,212-2 were purchased from Sigma-Aldrich (St. Louis, MO).

    Blocking Assay:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: .. After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents. .. The primary antibodies included anti-E-cadherin (1:1000, ab1416, Abcam), anti-Vimentin (1:1000, ab137321, Abcam), anti-β-actin (1:1000, ab227387, Abcam), and anti-Snail (1:1000, ab53519, Abcam).

    Polyacrylamide Gel Electrophoresis:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: The cell lysates (10–20 µg/lane) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on 10–12% gels and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, USA). .. After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents.

    Staining:

    Article Title: Primary cilia mediate mitochondrial stress responses to promote dopamine neuron survival in a Parkinson’s disease model
    Article Snippet: Paragraph title: Cilia staining and counting ... Subsequently, the cells were blocked with PBS containing 1% bovine serum albumin (BSA), and incubated overnight at 4 °C with primary antibodies against acetylated α-tubulin (1:1000, T7451, Sigma-Aldrich) or ARL13B (1:1000, 17711-1-AP, Proteintech) in 1% BSA.

    Article Title: In Vivo Effects of Ozone Exposure on Protein Adduct Formation by 1-Nitronaphthalene in Rat Lung
    Article Snippet: Bovine serum albumin (BSA), thiourea, tributylphosphine (TBP), and dithiothreitol (DTT) were ordered from Sigma-Aldrich (St. Louis, MO). .. Duracryl was obtained from Genomic Solutions (Ann Arbor, MI), and Rhinohide and SYPRO Ruby protein stain were purchased from Molecular Probes (Eugene, OR).

    Purification:

    Article Title: Initial synthesis and characterization of an immobilized heat shock protein 90 column for online determination of binding affinities
    Article Snippet: Bovine serum albumin (BSA), ammonium acetate, ATP, 1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC), glutaraldehyde, glutaric acid, glycine, pyridine (99.8%), sodium azide, and Tris buffer were obtained from Sigma-Aldrich Chemical Co. (St. Louis, MO, U.S.A.). .. The water used in the study was prepared using a Milli-QWater Purification System (Millipore Corporation, Bedford, MA, U.S.A.).

    SDS Page:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: The purity of the protein was > 96% and the molecular weight of the protein was approximately 175 kDa (confirmed with non-reducing SDS-PAGE and size-exclusion chromatography). .. Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: The cell lysates (10–20 µg/lane) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on 10–12% gels and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, USA). .. After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents.

    Proliferation Assay:

    Article Title: Preservation and Expansion of the Primate Keratocyte Phenotype by Downregulating TGF-? Signaling in a Low-Calcium, Serum-Free Medium
    Article Snippet: The tissue culture plastic plates (96-well and 6-well) were purchased from BD Biosciences (Lincoln Park, NJ); collagenase A and dispase II powder from Roche (Indianapolis, IN); amphotericin B, Dulbecco’s modified Eagle’s medium (DMEM), defined keratinocyte serum-free-medium (KSFM), FBS, gentamicin, Hanks ‘ balanced salt solution (HBSS), HEPES buffer, phosphate-buffered saline (PBS), soybean trypsin inhibitor, and 0.25% trypsin/1 mM EDTA were purchased from Invitrogen-Gibco (Grand Island, NY); A cell-viability–cytotoxicity kit (Live/Dead) from Invitrogen (Eugene, OR); endo-β-galactosidase was from Seikagaku (Tokyo, Japan); optimal cutting temperature (OCT) from Sakura Finetek (Torrance, CA); and a cell-proliferation assay (MTT) from Roche. .. Other reagents and chemicals including bovine serum albumin (BSA) and insulin-transferrin-sodium selenite (ITS) medium supplement were from Sigma-Aldrich.

    Software:

    Article Title: Evidence for the multimeric structure of ferroportin
    Article Snippet: Immunofluorescence was performed as previously described, with the following changes: after formaldehyde fixations, cells were incubated in phosphate-buffered saline (PBS), and 1% bovine serum albumin (BSA) with or without 0.1% saponin (Sigma, St Louis, MO). .. Images were captured on an Olympus BX51 microscope (Olympus, Tokyo, Japan) using an Olympus U-CMAD-Z camera and a 60×/1.3 NA oil objective with images acquired using Picture Frame 2.5 software.

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents. .. The relative levels of each protein expression were determined by densitometric analysis by using ImageJ software.

    Microscopy:

    Article Title: Primary cilia mediate mitochondrial stress responses to promote dopamine neuron survival in a Parkinson’s disease model
    Article Snippet: Subsequently, the cells were blocked with PBS containing 1% bovine serum albumin (BSA), and incubated overnight at 4 °C with primary antibodies against acetylated α-tubulin (1:1000, T7451, Sigma-Aldrich) or ARL13B (1:1000, 17711-1-AP, Proteintech) in 1% BSA. .. Cilia images were observed using a fluorescence microscope.

    Article Title: Evidence for the multimeric structure of ferroportin
    Article Snippet: Immunofluorescence was performed as previously described, with the following changes: after formaldehyde fixations, cells were incubated in phosphate-buffered saline (PBS), and 1% bovine serum albumin (BSA) with or without 0.1% saponin (Sigma, St Louis, MO). .. Images were captured on an Olympus BX51 microscope (Olympus, Tokyo, Japan) using an Olympus U-CMAD-Z camera and a 60×/1.3 NA oil objective with images acquired using Picture Frame 2.5 software.

    Next-Generation Sequencing:

    Article Title: Vesicular Glutamate and GABA Transporters Sort to Distinct Sets of Vesicles in a Population of Presynaptic Terminals
    Article Snippet: Components of Durcupan ACM, sodium azide (NaN3 ), diaminobenzidine (DAB), human serum albumin (HSA), bovine serum albumin (BSA), sodium ethylenediamine tetraacetate (EDTA), polyethylene glycol (PEG), Trizma base and Tris–HCl were obtained from Sigma Aldrich (Oslo, Norway). .. Newborn calf serum (NCS) and normal goat serum (NGS) were from Gibco (Paisley, UK).

    Produced:

    Article Title: Development of a Stable Respiratory Syncytial Virus Pre-Fusion Protein Powder Suitable for a Core-Shell Implant with a Delayed Release in Mice: A Proof of Concept Study
    Article Snippet: Materials Trimeric RSV pre-F protein (3.38 mg/mL (based on UV 280 nm) in 40 mM Tris buffer pH 7.5 with 150 mM NaCl), comparable to the vaccine candidate described by Krarup et al. [ ], was produced by U-Protein Express BV (Utrecht, The Netherlands) on request of the authors. .. Sodium phosphate dibasic, Tween 20, Tween 80, bovine serum albumin (BSA), Tris base, 8-anilino-1-napthalenesulfonic acid ammonium salt (ANS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and l -leucine were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).

    Concentration Assay:

    Article Title: Separation of the convulsions and antidepressant-like effects produced by the delta-opioid agonist SNC80 in rats
    Article Snippet: For standard curve determinations, 400 μl of vehicle-treated rat brains were added to 20 μl of a known concentration of SNC80 (0.032–10 μM) and 20 μl of 0.1 nM [3 H]NTI. .. Known concentrations of SNC80 diluted in 0.1% bovine serum albumin (BSA) were added to the assay tubes with Sigmacote-coated (Sigma, St. Louis, MO) pipette tips.

    Lysis:

    Article Title: CircRNA-5692 inhibits the progression of hepatocellular carcinoma by sponging miR-328-5p to enhance DAB2IP expression
    Article Snippet: Western blotting The different groups of cells were lysed in lysis buffer supplemented with protease inhibitors (Roche, Indianapolis, USA) and centrifuged. .. After blocking with 5% bovine serum albumin (BSA), the membranes were probed with primary antibodies for 4 h at 37 °C and detected with horseradish peroxidase (HRP)-conjugated secondary antibodies (1:5000, AP308P, Sigma-Aldrich), followed by visualization by using enhanced chemiluminescent reagents.

    Article Title: Characterization of the Early Steps of Hepatitis C Virus Infection by Using Luciferase Reporter Viruses
    Article Snippet: Huh7-Lunet cells were washed with PBS and then incubated with a buffer consisting of 20 mM Tris-HCl pH 6.8, 50 mM NaCl, 4 mM CaCl2 , 0.01% bovine serum albumin (BSA) (mock treated) or the same buffer containing 0.5 U/ml heparinase I (Sigma-Aldrich), or heparinase III (Sigma-Aldrich) or chondroitinase ABC (Sigma-Aldrich). .. After incubation for 1 h at 37°C, cells were washed three times with PBS and then inoculated with 500 μl of reporter virus preparation for 1 h. Forty-eight hours later, cells were lysed in 350 μl of lysis buffer, and luciferase activity was measured as described above.

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    Millipore bsa
    Penetration of C. neoformans through reconstituted ECM. The ECM invasion chambers are composed of matrigel (basement membrane) layered on membranes with 8 µm pores. Strain JEC21 was incubated with plasminogen in phosphate-buffered saline with <t>BSA</t> in the presence or absence of tissue-derived plasminogen activator (tPA), incubated in the upper chamber of the transwell for 24 hours at <t>37°C</t> prior to analysis of colony counts from the lower well (* (p = 0.0093); ** (p = 0.0084)).
    Bsa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1385 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Penetration of C. neoformans through reconstituted ECM. The ECM invasion chambers are composed of matrigel (basement membrane) layered on membranes with 8 µm pores. Strain JEC21 was incubated with plasminogen in phosphate-buffered saline with BSA in the presence or absence of tissue-derived plasminogen activator (tPA), incubated in the upper chamber of the transwell for 24 hours at 37°C prior to analysis of colony counts from the lower well (* (p = 0.0093); ** (p = 0.0084)).

    Journal: PLoS ONE

    Article Title: Surface-Associated Plasminogen Binding of Cryptococcus neoformans Promotes Extracellular Matrix Invasion

    doi: 10.1371/journal.pone.0005780

    Figure Lengend Snippet: Penetration of C. neoformans through reconstituted ECM. The ECM invasion chambers are composed of matrigel (basement membrane) layered on membranes with 8 µm pores. Strain JEC21 was incubated with plasminogen in phosphate-buffered saline with BSA in the presence or absence of tissue-derived plasminogen activator (tPA), incubated in the upper chamber of the transwell for 24 hours at 37°C prior to analysis of colony counts from the lower well (* (p = 0.0093); ** (p = 0.0084)).

    Article Snippet: Surface plasminogen activation Approximately 1×108 log phase cells were washed in PBS and incubated with 100 µg plasminogen for 2–4 hrs at 37°C in PBS with 1.5% BSA, in the presence or absence of 100 ng tissue plasminogen activator (tPA) (Calbiochem) or aprotinin (Roche).

    Techniques: Incubation, Derivative Assay

    Plasminogen binds selectively and specifically to the cell-surface of intact C. neoformans strains. (A–B) Conversion of plasminogen (Plg) into plasmin heavy chain (Pla H ) and light chain (Pla L ) on the surface of intact C. neoformans serotype D and A strains. (A) Serotype D strain JEC21 was incubated in the presence or absence of plasminogen, tissue plasminogen activator (tPA), and/or the plasmin inhibitor aprotinin in phosphate-buffered saline with BSA. Cell wall proteins were released by boiling labeled cells in SDS-extraction buffer and fractionated by SDS-PAGE, transferred to PVDF, and Western blotted with polyclonal anti-plasminogen antibody. Lane descriptions as follow: cells (JEC21) only (1), 100 µg plasminogen (2), plasminogen and 100 ng tPA (3), plasminogen, tPA, and 1 unit aprotinin (4). (B) Serotype A strains C23 and A1 38-2 were incubated in the presence or absence of plasminogen and/or tPA for 4 hrs at 37°C prior to Western blot analysis as described above. Lanes: cells (C23) only (1), C23 with 15 µg plasminogen (2), C23 with plasminogen and 100 ng tPA (3), cells (A1 38-2) only (4), A1 38-2 with 15 µg plasminogen (5), and A1 38-2 with plasminogen and tPA (6). (C) Plasminogen associates with the cell wall of intact cells. Cells (1×10 10 ) from log phase cultures (JEC21) were incubated 4 hr at 37°C in the presence (lane 1) or absence (lane 3) of 50 µg plasminogen and separated into cell wall and cytosol components, as described in Methods . Membranes (lane 2, 4) from cell walls were extracted and each fraction examined for the presence of plasminogen by Western blot analysis. Sample loading was uniform at 5 µg per well. (D) Sulfo-NHS-biotin and plasminogen compete for cell-surface binding sites. Log phase cells (JEC21) were initially labeled with sulfo-NHS-biotin in 0-, 1-, 10-, 100-fold molar equivalents of plasminogen then labeled 1 hr at 37°C with 50 µg plasminogen (lanes 1–4, respectively).

    Journal: PLoS ONE

    Article Title: Surface-Associated Plasminogen Binding of Cryptococcus neoformans Promotes Extracellular Matrix Invasion

    doi: 10.1371/journal.pone.0005780

    Figure Lengend Snippet: Plasminogen binds selectively and specifically to the cell-surface of intact C. neoformans strains. (A–B) Conversion of plasminogen (Plg) into plasmin heavy chain (Pla H ) and light chain (Pla L ) on the surface of intact C. neoformans serotype D and A strains. (A) Serotype D strain JEC21 was incubated in the presence or absence of plasminogen, tissue plasminogen activator (tPA), and/or the plasmin inhibitor aprotinin in phosphate-buffered saline with BSA. Cell wall proteins were released by boiling labeled cells in SDS-extraction buffer and fractionated by SDS-PAGE, transferred to PVDF, and Western blotted with polyclonal anti-plasminogen antibody. Lane descriptions as follow: cells (JEC21) only (1), 100 µg plasminogen (2), plasminogen and 100 ng tPA (3), plasminogen, tPA, and 1 unit aprotinin (4). (B) Serotype A strains C23 and A1 38-2 were incubated in the presence or absence of plasminogen and/or tPA for 4 hrs at 37°C prior to Western blot analysis as described above. Lanes: cells (C23) only (1), C23 with 15 µg plasminogen (2), C23 with plasminogen and 100 ng tPA (3), cells (A1 38-2) only (4), A1 38-2 with 15 µg plasminogen (5), and A1 38-2 with plasminogen and tPA (6). (C) Plasminogen associates with the cell wall of intact cells. Cells (1×10 10 ) from log phase cultures (JEC21) were incubated 4 hr at 37°C in the presence (lane 1) or absence (lane 3) of 50 µg plasminogen and separated into cell wall and cytosol components, as described in Methods . Membranes (lane 2, 4) from cell walls were extracted and each fraction examined for the presence of plasminogen by Western blot analysis. Sample loading was uniform at 5 µg per well. (D) Sulfo-NHS-biotin and plasminogen compete for cell-surface binding sites. Log phase cells (JEC21) were initially labeled with sulfo-NHS-biotin in 0-, 1-, 10-, 100-fold molar equivalents of plasminogen then labeled 1 hr at 37°C with 50 µg plasminogen (lanes 1–4, respectively).

    Article Snippet: Surface plasminogen activation Approximately 1×108 log phase cells were washed in PBS and incubated with 100 µg plasminogen for 2–4 hrs at 37°C in PBS with 1.5% BSA, in the presence or absence of 100 ng tissue plasminogen activator (tPA) (Calbiochem) or aprotinin (Roche).

    Techniques: Proximity Ligation Assay, Incubation, Labeling, SDS Page, Western Blot, Binding Assay

    Effects of bromodomain inhibitor in DN. MS417, a bromodomain inhibitor, suppressed acetylation of Stat3 and NF-κB in podocytes treated with AGE. Podocytes were incubated with either BSA or AGE-BSA together with DMSO or MS417 (1.0 μmol/L) for 24 h. Western blot analysis was performed in these cells for acetyl, phosphor- and total p65 ( A ) and STAT3 ( C ). The representative blots of three independent experiments are shown. The densitometry analyses of these Western blots are shown for p65 ( B ) and STAT3 ( D ). The ratios of acetyl-protein or phosphor-protein to total protein were calculated for p65 and STAT3. The fold changes relative to cells treated with BSA + DMSO are shown. * P

    Journal: Diabetes

    Article Title: Role of Transcription Factor Acetylation in Diabetic Kidney Disease

    doi: 10.2337/db13-1810

    Figure Lengend Snippet: Effects of bromodomain inhibitor in DN. MS417, a bromodomain inhibitor, suppressed acetylation of Stat3 and NF-κB in podocytes treated with AGE. Podocytes were incubated with either BSA or AGE-BSA together with DMSO or MS417 (1.0 μmol/L) for 24 h. Western blot analysis was performed in these cells for acetyl, phosphor- and total p65 ( A ) and STAT3 ( C ). The representative blots of three independent experiments are shown. The densitometry analyses of these Western blots are shown for p65 ( B ) and STAT3 ( D ). The ratios of acetyl-protein or phosphor-protein to total protein were calculated for p65 and STAT3. The fold changes relative to cells treated with BSA + DMSO are shown. * P

    Article Snippet: Chromatin Immunoprecipitation Assay Human podocytes were infected with the following viruses: green fluorescent protein, wild-type and K658R STAT3 or green fluorescent protein, wild-type and K310R p65 (1:100 V/V) for 3 days and then treated with either AGE-BSA (100 μg/mL) or BSA (100 μg/mL) for an additional 24 h. Chromatin immunoprecipitation assay was performed according to the EZ-CHIP protocol (Millipore catalog no. 17–371) ( ).

    Techniques: Incubation, Western Blot

    AGE induced acetylation of p65 and STAT3, which is required for their transcriptional activation. A : Human podocytes were cultured with AGE or BSA at the indicated doses for 24 h. Cell lysates were subject to Western blot analysis for acetyl-, phosphor-, and total p65 and STAT3. The representative blots of three independent experiments are shown. B : The Western blots from all experiments were quantified by densitometry analysis. The ratios of acetyl-protein or phosphor-protein to total protein were calculated for p65 and STAT3. The fold changes relative to BSA-treated cells are shown. * P

    Journal: Diabetes

    Article Title: Role of Transcription Factor Acetylation in Diabetic Kidney Disease

    doi: 10.2337/db13-1810

    Figure Lengend Snippet: AGE induced acetylation of p65 and STAT3, which is required for their transcriptional activation. A : Human podocytes were cultured with AGE or BSA at the indicated doses for 24 h. Cell lysates were subject to Western blot analysis for acetyl-, phosphor-, and total p65 and STAT3. The representative blots of three independent experiments are shown. B : The Western blots from all experiments were quantified by densitometry analysis. The ratios of acetyl-protein or phosphor-protein to total protein were calculated for p65 and STAT3. The fold changes relative to BSA-treated cells are shown. * P

    Article Snippet: Chromatin Immunoprecipitation Assay Human podocytes were infected with the following viruses: green fluorescent protein, wild-type and K658R STAT3 or green fluorescent protein, wild-type and K310R p65 (1:100 V/V) for 3 days and then treated with either AGE-BSA (100 μg/mL) or BSA (100 μg/mL) for an additional 24 h. Chromatin immunoprecipitation assay was performed according to the EZ-CHIP protocol (Millipore catalog no. 17–371) ( ).

    Techniques: Activation Assay, Cell Culture, Western Blot

    Acetylation of p65 and STAT3 is required for AGE-induced transcriptional activation of their target genes. Podocytes were infected with p65 or STAT3 constructs with mutated acetyl residues for 3 days then stimulated with AGE or BSA at the indicated doses for an additional 24 h. Total RNA was isolated from these cells for real-time PCR analysis of NF-κB target genes ( CXCL5 [ A ] and CCL17 [ B ]) and STAT3 target genes ( SOC3 [ C ] and Casp9 [ D ]). For chromatin immunoprecipitation (ChiP) assay for NF-κB and STAT3 target genes, podocytes were infected with p65 or STAT3 constructs with mutated acetyl residues for 3 days then stimulated with AGE or BSA at the indicated doses for an additional 24 h. ChiP assay also was performed in these cells to determine the binding of p65 and STAT3 on the promoter sites of their target genes ( CXCL5 [ E ], CCL17 [ F ], SOC3 [ G ], and Casp9 [ H ]). Immunoprecipitated DNA samples were subjected to PCR analysis and are expressed as a percentage of input. * P

    Journal: Diabetes

    Article Title: Role of Transcription Factor Acetylation in Diabetic Kidney Disease

    doi: 10.2337/db13-1810

    Figure Lengend Snippet: Acetylation of p65 and STAT3 is required for AGE-induced transcriptional activation of their target genes. Podocytes were infected with p65 or STAT3 constructs with mutated acetyl residues for 3 days then stimulated with AGE or BSA at the indicated doses for an additional 24 h. Total RNA was isolated from these cells for real-time PCR analysis of NF-κB target genes ( CXCL5 [ A ] and CCL17 [ B ]) and STAT3 target genes ( SOC3 [ C ] and Casp9 [ D ]). For chromatin immunoprecipitation (ChiP) assay for NF-κB and STAT3 target genes, podocytes were infected with p65 or STAT3 constructs with mutated acetyl residues for 3 days then stimulated with AGE or BSA at the indicated doses for an additional 24 h. ChiP assay also was performed in these cells to determine the binding of p65 and STAT3 on the promoter sites of their target genes ( CXCL5 [ E ], CCL17 [ F ], SOC3 [ G ], and Casp9 [ H ]). Immunoprecipitated DNA samples were subjected to PCR analysis and are expressed as a percentage of input. * P

    Article Snippet: Chromatin Immunoprecipitation Assay Human podocytes were infected with the following viruses: green fluorescent protein, wild-type and K658R STAT3 or green fluorescent protein, wild-type and K310R p65 (1:100 V/V) for 3 days and then treated with either AGE-BSA (100 μg/mL) or BSA (100 μg/mL) for an additional 24 h. Chromatin immunoprecipitation assay was performed according to the EZ-CHIP protocol (Millipore catalog no. 17–371) ( ).

    Techniques: Activation Assay, Infection, Construct, Isolation, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Binding Assay, Immunoprecipitation, Polymerase Chain Reaction

    Sca-1 repression leads to increased adhesion to collagen I, collagen IV and fibronectin. Cell adhesion of 100,000 cells/well coated with fibronectin, vitronectin, laminin, collagen I or collagen IV comparing shLuc (open bars), shSca-1 (grey bars) and shSca-1+Sca-1 (black bars). (A). Relative adhesion was normalized to a BSA control, (* p

    Journal: PLoS ONE

    Article Title: Stem Cell Antigen-1 (Sca-1) Regulates Mammary Tumor Development and Cell Migration

    doi: 10.1371/journal.pone.0027841

    Figure Lengend Snippet: Sca-1 repression leads to increased adhesion to collagen I, collagen IV and fibronectin. Cell adhesion of 100,000 cells/well coated with fibronectin, vitronectin, laminin, collagen I or collagen IV comparing shLuc (open bars), shSca-1 (grey bars) and shSca-1+Sca-1 (black bars). (A). Relative adhesion was normalized to a BSA control, (* p

    Article Snippet: Adhesion Assay 105 cells were seeded onto CytoMatrix™ Cell Adhesion Strips coated with BSA, Collagen Type I and IV, Fibronectin, Laminin, Vitronectin (Millipore) according to the manufacturer's guidelines.

    Techniques:

    Overexpression of ADAM10 promotes proliferation and migration of HASMCs in low glucose, high glucose and high glucose medium with addition of AGE-BSA (100 and 200 µg/ml). A , vector-transduced, ADAM10-overexpressing and ADAM10-silenced HASMCs were generated by retrovirus-mediated gene transfer and selection. These HASMCs were grown in low glucose, high glucose and high glucose with addition of AGE-BSA (100 and 200 ug/ml). MTT assay was performed to test the cell viability. *P

    Journal: PLoS ONE

    Article Title: Increase of ADAM10 Level in Coronary Artery In-Stent Restenosis Segments in Diabetic Minipigs: High ADAM10 Expression Promoting Growth and Migration in Human Vascular Smooth Muscle Cells via Notch 1 and 3

    doi: 10.1371/journal.pone.0083853

    Figure Lengend Snippet: Overexpression of ADAM10 promotes proliferation and migration of HASMCs in low glucose, high glucose and high glucose medium with addition of AGE-BSA (100 and 200 µg/ml). A , vector-transduced, ADAM10-overexpressing and ADAM10-silenced HASMCs were generated by retrovirus-mediated gene transfer and selection. These HASMCs were grown in low glucose, high glucose and high glucose with addition of AGE-BSA (100 and 200 ug/ml). MTT assay was performed to test the cell viability. *P

    Article Snippet: AGE-BSA was from Calbiochem (Merck, Darmstadt, Germany).

    Techniques: Over Expression, Migration, Plasmid Preparation, Generated, Selection, MTT Assay