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Bio-Rad ethylene glycol tetraacetic acid egta
Ethylene Glycol Tetraacetic Acid Egta, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 1 article reviews
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ethylene glycol tetraacetic acid egta - by Bioz Stars, 2020-07
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Lysis:

Article Title: Dracorhodin Perchlorate Induced Human Breast Cancer MCF-7 Apoptosis through Mitochondrial Pathways
Article Snippet: .. Briefly, the cell pellets were resuspended in lysis buffer, including 50 mmol/L HEPES (pH 7.4), 1% Triton-X 100, 2 mmol/L sodium orthovanadate, 100 mmol/L sodium fluoride, 1 mmol/L EDTA, 1 mmol/L egtazic acid (EGTA), 1 mmol/L phenylmethylsulfonyl fluoride (PMSF), 0.1 g/L aprotinin, and 0.01 g/L leupeptin, then lysed at 4 °C for 1 h. After 13 000×g centrifugation for 10 min, the protein content of the supernatant was determined using Bio-Rad protein assay reagent (Bio-Rad, USA). .. The protein was loaded in each lane, then separated by 12% SDS-PAGE, and blotted onto a nitrocellulose membrane.

Article Title: Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells
Article Snippet: .. The cell pellets were resuspended in lysis buffer containing 50 mmol/L Hepes (pH 7.4), 1% Triton-X 100, 2 mmol/L sodium orthovanadate, 100 mmol/L sodium fluoride (NaF), 1 mmol/L edetic acid, 1 mmol/L egtazic acid (EGTA), 1 mmol/L phenylmethyl-sulfonylfluoride (PMSF), 0.1 g/L aprotinin, and 0.01 g/L leupeptin and lysed at 4 o C for 1 h. Then the cells were spun in a centrifuge at 12 000×g for 10 min, and the protein content of the supernatant was determined by Bio-Rad protein assay reagent (Bio-Rad, Hercules, CA, USA). .. The proteins were separated by 12% SDS polyacrylamide gel electrophoresis and blotted onto nitrocellulose membrane.

Article Title: Pseudolaric Acid B Induced Cell Cycle Arrest, Autophagy and Senescence in Murine Fibrosarcoma L929 Cell
Article Snippet: .. Briefly, the cell pellets were resuspended in lysis buffer, including Hepes 50 mmol/L pH 7.4, 1% Triton-X 100, sodium orthovanadate 2 mmol/L, sodium fluoride (NaF) 100 mmol/L, edetic acid 1 mmol/L, egtazic acid (EGTA) 1 mmol/L, phenylmethyl-sulfonylfluoride (PMSF) 1 mmol/L, aprotinin 0.1 g/L, leupeptin 0.01 g/L, then lysed in 4 o C for 1 h. After 13,000 × g centrifugation for 10 min, the protein content of the supernatant was determined using Bio-Rad protein assay reagent (Bio-Rad, Hercules, CA, USA). ..

Article Title: A dominant connexin43 mutant does not have dominant effects on gap junction coupling in astrocytes
Article Snippet: .. Individual brains, cerebella, cervical spinal cords and hearts were obtained from euthanized P44 Gja1Jrt/ + and their Gja1 + / + littermates, homogenized in lysis buffer (150 mM NaCl, 10 mM Tris, 1 mM ethylene diamine tetraacetic acid (EDTA), 1 mM ethylene glycol tetraacetic acid (EGTA), 1% Triton X, 1 mM Na vanadate and 1 mM NaFl; ) and separated on 4–15% Tris–HCL ReadyGel (Bio-Rad). .. The blots were incubated overnight at 4°C with a rabbit antiserum against Cx43 (1:8000 or 1:80,000 dilution; Sigma) or Cx30 (1:500 dilution; Zymed), washed several times, incubated in horse radish peroxidase (HRP)-conjugated donkey anti-rabbit antiserum (1:10,000 dilution; Jackson ImmunoResearch Laboratories) and developed using Amersham ECL Western Blotting Detection Reagents.

Article Title: Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells
Article Snippet: .. The cell pellets were resuspended in lysis buffer containing 50 mmol/L Hepes (pH 7.4), 1% Triton-X 100, 2 mmol/L sodium orthovanadate, 100 mmol/L sodium fluoride (NaF), 1 mmol/L edetic acid, 1 mmol/L egtazic acid (EGTA), 1 mmol/L phenylmethyl-sulfonylfluoride (PMSF), 0.1 g/L aprotinin, and 0.01 g/L leupeptin and lysed at 4 o C for 1 h. Then the cells were spun in a centrifuge at 12 000× g for 10 min, and the protein content of the supernatant was determined by Bio-Rad protein assay reagent (Bio-Rad, Hercules, CA, USA). .. The proteins were separated by 12% SDS polyacrylamide gel electrophoresis and blotted onto nitrocellulose membrane.

Centrifugation:

Article Title: Dracorhodin Perchlorate Induced Human Breast Cancer MCF-7 Apoptosis through Mitochondrial Pathways
Article Snippet: .. Briefly, the cell pellets were resuspended in lysis buffer, including 50 mmol/L HEPES (pH 7.4), 1% Triton-X 100, 2 mmol/L sodium orthovanadate, 100 mmol/L sodium fluoride, 1 mmol/L EDTA, 1 mmol/L egtazic acid (EGTA), 1 mmol/L phenylmethylsulfonyl fluoride (PMSF), 0.1 g/L aprotinin, and 0.01 g/L leupeptin, then lysed at 4 °C for 1 h. After 13 000×g centrifugation for 10 min, the protein content of the supernatant was determined using Bio-Rad protein assay reagent (Bio-Rad, USA). .. The protein was loaded in each lane, then separated by 12% SDS-PAGE, and blotted onto a nitrocellulose membrane.

Article Title: Pseudolaric Acid B Induced Cell Cycle Arrest, Autophagy and Senescence in Murine Fibrosarcoma L929 Cell
Article Snippet: .. Briefly, the cell pellets were resuspended in lysis buffer, including Hepes 50 mmol/L pH 7.4, 1% Triton-X 100, sodium orthovanadate 2 mmol/L, sodium fluoride (NaF) 100 mmol/L, edetic acid 1 mmol/L, egtazic acid (EGTA) 1 mmol/L, phenylmethyl-sulfonylfluoride (PMSF) 1 mmol/L, aprotinin 0.1 g/L, leupeptin 0.01 g/L, then lysed in 4 o C for 1 h. After 13,000 × g centrifugation for 10 min, the protein content of the supernatant was determined using Bio-Rad protein assay reagent (Bio-Rad, Hercules, CA, USA). ..

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  • 88
    Bio-Rad dntb egta
    Vespula spp and Apis mellifera venom PLA2 activity. (A) PLA2 activity was detected as free thiol groups released from thiol-labeled lipid substrates through addition of <t>DNTB/EGTA</t> and measuring color generation at 415 nm using Apis mellifera venom with four phosphatidylcholine-based substrates of varying acyl chain lengths and saturation, but the same headgroup. Vespula spp venom PLA2 activity for substrates diheptanoyl-phosphatidylcholine and arachidonyl-phosphatidylcholine (P
    Dntb Egta, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dntb egta/product/Bio-Rad
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dntb egta - by Bioz Stars, 2020-07
    88/100 stars
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    85
    Bio-Rad immunoblot analysis homogenate
    Function and cell-surface expression of DAT is reduced by synucleins. (A) Uptake of [ 3 H]-DA into SH-SY5Y cells co-transfected with DAT (100 ng/cm 2 ) and increasing amounts (100–400 ng/cm 2 ) of β-Syn (orange open circles) or γ-Syn (red open squares) was measured over 10 min. Values (n = 6; mean ± SEM) are presented as percent of DAT alone (line at 100%). Non-specific uptake in the presence of 10 μM indatraline was subtracted. As a positive control for Syn modulation of DAT, uptake was also measured in cells co-transfected with DAT and 400 ng/cm 2 of α-Syn (yellow triangle). Total amount of transfected DNA was kept constant at 500 ng/cm 2 with the addition of empty pcDNA3.1 vector. (B) Expression levels of α-Syn, β-Syn, γ-Syn, and DAT in cells used for uptake assays were confirmed by <t>immunoblot</t> (IB). Representative blot images are shown and quantified relative to actin in the adjacent graphs (n = 3; mean ± SEM). (C) Viability was assessed by MTT assay in SH-SY5Y cells transfected under conditions identical to the uptake experiment. Results from three independent experiments assayed in quadruplicate are expressed as absorbance (OD) at 570 nm ± SEM. (D) Cell surface protein was biotinylated in SH-SY5Y cells co-transfected at the 4∶1 ratio as above. Biotinylated DAT (IB DAT Biotin) captured with streptavidin beads and total DAT (IB DAT Total) were measured by immunoblot. DAT biotinylation was quantified as optical density (OD) of DAT Biotin divided by OD of DAT Total relative to actin (n = 4; mean ± SEM). Molecular mass (M r ) of nearest protein ladder bands is indicated. Data in were analyzed by t-test for difference from a theoretical mean of 100 (*p
    Immunoblot Analysis Homogenate, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoblot analysis homogenate/product/Bio-Rad
    Average 85 stars, based on 1 article reviews
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    88
    Bio-Rad cytomix buffer
    Function and cell-surface expression of DAT is reduced by synucleins. (A) Uptake of [ 3 H]-DA into SH-SY5Y cells co-transfected with DAT (100 ng/cm 2 ) and increasing amounts (100–400 ng/cm 2 ) of β-Syn (orange open circles) or γ-Syn (red open squares) was measured over 10 min. Values (n = 6; mean ± SEM) are presented as percent of DAT alone (line at 100%). Non-specific uptake in the presence of 10 μM indatraline was subtracted. As a positive control for Syn modulation of DAT, uptake was also measured in cells co-transfected with DAT and 400 ng/cm 2 of α-Syn (yellow triangle). Total amount of transfected DNA was kept constant at 500 ng/cm 2 with the addition of empty pcDNA3.1 vector. (B) Expression levels of α-Syn, β-Syn, γ-Syn, and DAT in cells used for uptake assays were confirmed by <t>immunoblot</t> (IB). Representative blot images are shown and quantified relative to actin in the adjacent graphs (n = 3; mean ± SEM). (C) Viability was assessed by MTT assay in SH-SY5Y cells transfected under conditions identical to the uptake experiment. Results from three independent experiments assayed in quadruplicate are expressed as absorbance (OD) at 570 nm ± SEM. (D) Cell surface protein was biotinylated in SH-SY5Y cells co-transfected at the 4∶1 ratio as above. Biotinylated DAT (IB DAT Biotin) captured with streptavidin beads and total DAT (IB DAT Total) were measured by immunoblot. DAT biotinylation was quantified as optical density (OD) of DAT Biotin divided by OD of DAT Total relative to actin (n = 4; mean ± SEM). Molecular mass (M r ) of nearest protein ladder bands is indicated. Data in were analyzed by t-test for difference from a theoretical mean of 100 (*p
    Cytomix Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cytomix buffer/product/Bio-Rad
    Average 88 stars, based on 18 article reviews
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    93
    Bio-Rad egta
    Function and cell-surface expression of DAT is reduced by synucleins. (A) Uptake of [ 3 H]-DA into SH-SY5Y cells co-transfected with DAT (100 ng/cm 2 ) and increasing amounts (100–400 ng/cm 2 ) of β-Syn (orange open circles) or γ-Syn (red open squares) was measured over 10 min. Values (n = 6; mean ± SEM) are presented as percent of DAT alone (line at 100%). Non-specific uptake in the presence of 10 μM indatraline was subtracted. As a positive control for Syn modulation of DAT, uptake was also measured in cells co-transfected with DAT and 400 ng/cm 2 of α-Syn (yellow triangle). Total amount of transfected DNA was kept constant at 500 ng/cm 2 with the addition of empty pcDNA3.1 vector. (B) Expression levels of α-Syn, β-Syn, γ-Syn, and DAT in cells used for uptake assays were confirmed by <t>immunoblot</t> (IB). Representative blot images are shown and quantified relative to actin in the adjacent graphs (n = 3; mean ± SEM). (C) Viability was assessed by MTT assay in SH-SY5Y cells transfected under conditions identical to the uptake experiment. Results from three independent experiments assayed in quadruplicate are expressed as absorbance (OD) at 570 nm ± SEM. (D) Cell surface protein was biotinylated in SH-SY5Y cells co-transfected at the 4∶1 ratio as above. Biotinylated DAT (IB DAT Biotin) captured with streptavidin beads and total DAT (IB DAT Total) were measured by immunoblot. DAT biotinylation was quantified as optical density (OD) of DAT Biotin divided by OD of DAT Total relative to actin (n = 4; mean ± SEM). Molecular mass (M r ) of nearest protein ladder bands is indicated. Data in were analyzed by t-test for difference from a theoretical mean of 100 (*p
    Egta, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Bio-Rad
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    Image Search Results


    Vespula spp and Apis mellifera venom PLA2 activity. (A) PLA2 activity was detected as free thiol groups released from thiol-labeled lipid substrates through addition of DNTB/EGTA and measuring color generation at 415 nm using Apis mellifera venom with four phosphatidylcholine-based substrates of varying acyl chain lengths and saturation, but the same headgroup. Vespula spp venom PLA2 activity for substrates diheptanoyl-phosphatidylcholine and arachidonyl-phosphatidylcholine (P

    Journal: The Journal of Experimental Medicine

    Article Title: Bee venom processes human skin lipids for presentation by CD1a

    doi: 10.1084/jem.20141505

    Figure Lengend Snippet: Vespula spp and Apis mellifera venom PLA2 activity. (A) PLA2 activity was detected as free thiol groups released from thiol-labeled lipid substrates through addition of DNTB/EGTA and measuring color generation at 415 nm using Apis mellifera venom with four phosphatidylcholine-based substrates of varying acyl chain lengths and saturation, but the same headgroup. Vespula spp venom PLA2 activity for substrates diheptanoyl-phosphatidylcholine and arachidonyl-phosphatidylcholine (P

    Article Snippet: After 1 h incubation, 10 µl DNTB/EGTA was added, which reacts with free thiol groups producing a colored precipitate, which was measured at 415 nm wavelength (iMark Microplate Reader; Bio-Rad Laboratories).

    Techniques: Activity Assay, Labeling

    Function and cell-surface expression of DAT is reduced by synucleins. (A) Uptake of [ 3 H]-DA into SH-SY5Y cells co-transfected with DAT (100 ng/cm 2 ) and increasing amounts (100–400 ng/cm 2 ) of β-Syn (orange open circles) or γ-Syn (red open squares) was measured over 10 min. Values (n = 6; mean ± SEM) are presented as percent of DAT alone (line at 100%). Non-specific uptake in the presence of 10 μM indatraline was subtracted. As a positive control for Syn modulation of DAT, uptake was also measured in cells co-transfected with DAT and 400 ng/cm 2 of α-Syn (yellow triangle). Total amount of transfected DNA was kept constant at 500 ng/cm 2 with the addition of empty pcDNA3.1 vector. (B) Expression levels of α-Syn, β-Syn, γ-Syn, and DAT in cells used for uptake assays were confirmed by immunoblot (IB). Representative blot images are shown and quantified relative to actin in the adjacent graphs (n = 3; mean ± SEM). (C) Viability was assessed by MTT assay in SH-SY5Y cells transfected under conditions identical to the uptake experiment. Results from three independent experiments assayed in quadruplicate are expressed as absorbance (OD) at 570 nm ± SEM. (D) Cell surface protein was biotinylated in SH-SY5Y cells co-transfected at the 4∶1 ratio as above. Biotinylated DAT (IB DAT Biotin) captured with streptavidin beads and total DAT (IB DAT Total) were measured by immunoblot. DAT biotinylation was quantified as optical density (OD) of DAT Biotin divided by OD of DAT Total relative to actin (n = 4; mean ± SEM). Molecular mass (M r ) of nearest protein ladder bands is indicated. Data in were analyzed by t-test for difference from a theoretical mean of 100 (*p

    Journal: PLoS ONE

    Article Title: Synucleins Antagonize Endoplasmic Reticulum Function to Modulate Dopamine Transporter Trafficking

    doi: 10.1371/journal.pone.0070872

    Figure Lengend Snippet: Function and cell-surface expression of DAT is reduced by synucleins. (A) Uptake of [ 3 H]-DA into SH-SY5Y cells co-transfected with DAT (100 ng/cm 2 ) and increasing amounts (100–400 ng/cm 2 ) of β-Syn (orange open circles) or γ-Syn (red open squares) was measured over 10 min. Values (n = 6; mean ± SEM) are presented as percent of DAT alone (line at 100%). Non-specific uptake in the presence of 10 μM indatraline was subtracted. As a positive control for Syn modulation of DAT, uptake was also measured in cells co-transfected with DAT and 400 ng/cm 2 of α-Syn (yellow triangle). Total amount of transfected DNA was kept constant at 500 ng/cm 2 with the addition of empty pcDNA3.1 vector. (B) Expression levels of α-Syn, β-Syn, γ-Syn, and DAT in cells used for uptake assays were confirmed by immunoblot (IB). Representative blot images are shown and quantified relative to actin in the adjacent graphs (n = 3; mean ± SEM). (C) Viability was assessed by MTT assay in SH-SY5Y cells transfected under conditions identical to the uptake experiment. Results from three independent experiments assayed in quadruplicate are expressed as absorbance (OD) at 570 nm ± SEM. (D) Cell surface protein was biotinylated in SH-SY5Y cells co-transfected at the 4∶1 ratio as above. Biotinylated DAT (IB DAT Biotin) captured with streptavidin beads and total DAT (IB DAT Total) were measured by immunoblot. DAT biotinylation was quantified as optical density (OD) of DAT Biotin divided by OD of DAT Total relative to actin (n = 4; mean ± SEM). Molecular mass (M r ) of nearest protein ladder bands is indicated. Data in were analyzed by t-test for difference from a theoretical mean of 100 (*p

    Article Snippet: Protein extraction, SDS-PAGE and immunoblot analysis Homogenate (prepared in 10 mM Tris HCl, 100 mM NaCl, 1 mM EDTA, 1 mM EGTA, 250 mM sucrose, pH 7.4) protein concentration was determined by Lowry assay (Biorad), adjusted, and protein was extracted with the addition of detergents (1% Triton X-100, 0.5% Na-deoxycholate, 0.1% SDS) for 1 h at 4°C.

    Techniques: Expressing, Transfection, Positive Control, Plasmid Preparation, MTT Assay