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denaturing loading buffer  (Bio-Rad)


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    Bio-Rad denaturing loading buffer
    Denaturing Loading Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1967 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/denaturing+loading+buffer/Bromophenol+Blue/pmc12988328-148-6-44
    Average 96 stars, based on 1967 article reviews
    denaturing loading buffer - by Bioz Stars, 2026-10
    96/100 stars

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    Related Articles

    SDS Page:

    Article Title: Apolipoprotein D deficiency is associated to high bone turnover, low bone mass and impaired osteoblastic function in aged female mice
    Article Snippet: .. All samples were mixed with 5 × denaturing loading buffer and resolved on a 12% SDS-PAGE (Bio-Rad Laboratories, Mississauga, Canada). .. Gels were transferred onto PVDF membranes and immunoblotted with a mouse-specific goat anti-ApoD (1:1000, sc34760, Santa- Cruz sc34760 biotechnology, Dallas, TX, USA), followed by an anti-goat secondary antibody (1:10,000, Santa Cruz) coupled to HRP.

    Article Title: Reduced mitochondrial pyruvate carrier expression in hearts with heart failure and reduced ejection fraction patients: ischemic vs. non-ischemic origin
    Article Snippet: St. Louis, MO, USA), and quantified using a Pierce TM BCA Protein Assay Kit (Thermo Scientific, Rockford, IL, USA), following the manufacturer's instructions. .. After extracting protein from each tissue, 30 μg of protein was loaded into each lane of a 4%–12% gradient acrylamide Bio-Rad gel using denaturing loading buffer and proteins were separated by SDS-PAGE using the Mini-Protean 3 system (Bio-Rad, Madrid, Spain), followed by transfer to PVDF membranes (Bio-Rad). .. Blots were incubated with the following primary antibodies diluted in PBS-Tween 5% milk overnight at 4°C: MPC1 (Cell Signaling, #14462, Danvers, MA, USA), MPC2 (Cell Signaling, #14462), PDH (Abcam, ab110416, Cambridge, UK) and SIRT3 (Santa Cruz Biotechnology, ab1104169, Santa Cruz, CA, USA).

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 °C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 °C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic.
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 ◦C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 ◦C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.

    DC Protein Assay:

    Article Title: Development of ER-alpha and ER-beta expression in the developing ovine brain and pituitary.
    Article Snippet: Fetal neuroendocrine development in late gestation is critical for maintenance of fetal homeostasis, growth, and readiness for birth.. We designed the present study to identify the regional patterns of expression of the two main isoforms of the estrogen receptor, ER-a and ER-b, in the developing ovine fetal brain.. Fetal (80, 100, 120, 130, and 145 days gestation), neonatal (1 and 7 days), and adult sheep were euthanized and the following tissues were collected: pituitary, hypothalamus, hippocampus, cerebral cortex, and brainstem.

    Molecular Weight:

    Article Title: Development and validation of a new monoclonal antibody to mammalian aromatase.
    Article Snippet: Microsomal pellets were resuspended in 200 μl solubilisation buffer (50 mM potassium phosphate buffer, pH 7·4) containing 20% glycerol (v/v), 0·5% (w/v) sodium cholate (Sigma), 0·1 mM EDTA (Sigma), 1 mM DTT (Sigma), 0·2% (w/v) Igepal CO-990 (Aldrich, Gillingham, Dorset, UK), containing two protease inhibitor tablets/50 ml buffer). .. Aliquots (50 μg total protein) were added to 5 μl denaturing/ loading buffer (50 mM Tris–HCl, pH 6·8, 100 mM DTT, 2% SDS, 0·1% bromophenol blue, 10% glycerol, all from Sigma) and boiled for 5 min. Tissue extracts and prestained protein molecular weight markers (BioRad Laboratories Ltd, Hemel Hempstead, Herts, UK) were separated on denaturing minigels containing an acrylamide gradient from 4 to 20% (w/v) polyacrylamide (Invitrogen) run in www.endocrinology.org Journal of Endocrinology (2002) 172, 21–30 www.endocrinology.orgJournal of Endocrinology (2002) 172, 21–30 www.endocrinology.org Journal of Endocrinology (2002) 172, 21–30 an Xcell-II minicell tank using Tris/glycine/SDS running buffer (25 mM Tris, 250 mM glycine, pH 8·3, 0·1% SDS) at 100 V per gel for 2 h and blotted onto polyvinylidene difluoride membrane (Millipore, Watford, Herts, UK). ..

    Membrane:

    Article Title: Development and validation of a new monoclonal antibody to mammalian aromatase.
    Article Snippet: Microsomal pellets were resuspended in 200 μl solubilisation buffer (50 mM potassium phosphate buffer, pH 7·4) containing 20% glycerol (v/v), 0·5% (w/v) sodium cholate (Sigma), 0·1 mM EDTA (Sigma), 1 mM DTT (Sigma), 0·2% (w/v) Igepal CO-990 (Aldrich, Gillingham, Dorset, UK), containing two protease inhibitor tablets/50 ml buffer). .. Aliquots (50 μg total protein) were added to 5 μl denaturing/ loading buffer (50 mM Tris–HCl, pH 6·8, 100 mM DTT, 2% SDS, 0·1% bromophenol blue, 10% glycerol, all from Sigma) and boiled for 5 min. Tissue extracts and prestained protein molecular weight markers (BioRad Laboratories Ltd, Hemel Hempstead, Herts, UK) were separated on denaturing minigels containing an acrylamide gradient from 4 to 20% (w/v) polyacrylamide (Invitrogen) run in www.endocrinology.org Journal of Endocrinology (2002) 172, 21–30 www.endocrinology.orgJournal of Endocrinology (2002) 172, 21–30 www.endocrinology.org Journal of Endocrinology (2002) 172, 21–30 an Xcell-II minicell tank using Tris/glycine/SDS running buffer (25 mM Tris, 250 mM glycine, pH 8·3, 0·1% SDS) at 100 V per gel for 2 h and blotted onto polyvinylidene difluoride membrane (Millipore, Watford, Herts, UK). ..

    Radio Immunoprecipitation:

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 °C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 °C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic.
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 ◦C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 ◦C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.

    Incubation:

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 °C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 °C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.

    Article Title: Characterization of the First SARS-CoV-2 Isolates from Aotearoa New Zealand as Part of a Rapid Response to the COVID-19 Pandemic.
    Article Snippet: Cells were washed four times (5 min each) with PBS-T and images recorded with an Olympus IX71 inverted fluorescence microscope (Olympus, Tokio, Japan) using cellSens software (Olympus). .. Aliquots of the SARS-CoV-2 isolates (C1 viral stocks) were mixed 1:1 with radioimmunoprecipitation assay (RIPA) buffer (10 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 50 mM Tris pH 8.0, and 20 mM Tris-HCl pH 7.5), incubated for 10 min at room temperature, then frozen at −80 ◦C for 24 h. Protein lysates were thawed and mixed with 6× denaturing loading buffer (4% SDS, 5% β-mercaptoethanol, 20% glycerol, 0.004% bromophenol blue, and 0.125 M Tris HCl pH 6.8), heated at 70 ◦C for 15 min, loaded onto a 8% SDS-polyacrylamide gel topped with a 5% stacking gel (BioRad, Hercules, CA, USA), and run at 120 V for one hour in 1× SDS-PAGE running buffer. .. Proteins were transferred onto a Immobilon-FL membrane (Merck KGA, Darmstadt, Germany) in 1× transfer buffer (25 mM Tris base, 0.1% SDS, and 10% methanol) overnight at 15 V. Membranes were blocked in 0.5% BSA and 0.01% Tween-20 in PBS for one hour, then incubated with the primary antibody (SARS-CoV-2 Nucleocapsid Antibody, mouse mAb, ProSci Inc, Poway, CA, USA) in blocking solution for 2 h. Membranes were washed (3×) in PBS with 0.1% Tween 20, then incubated for one hour with the secondary antibody (Goat anti-mouse Alexa FluorTM Plus 800, Thermo Fisher Scientific) diluted 1:10,000 in blocking solution.



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