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Amersham protran premium nitrocellulose western blotting membranes

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Amersham Protran Premium Nitrocellulose Western Blotting Membranes are high-quality nitrocellulose membranes designed for western blotting applications. They provide consistent protein transfer and binding for effective protein detection and analysis.

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Lab products found in correlation

Mitochondria isolation kit, by Thermo Fisher Scientific (2 mentions) Novex empty gel cassettes, by Thermo Fisher Scientific (2 mentions) Nc3515, by Thermo Fisher Scientific (2 mentions) Agarose, by Bio-Rad (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Bis tris gel, by Thermo Fisher Scientific (2 mentions) Amersham ecl hrp, by GE Healthcare (2 mentions) Immobilon forte western hrp substrate, by Merck Group (2 mentions)

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4 protocols using amersham protran premium nitrocellulose western blotting membranes

1

In vivo MAVS Aggregation Assay

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In vivo MAVS aggregation assay was performed according to a published protocol (Hou et al., 2011 (link)). Briefly, crude mitochondria isolated using mitochondria isolation kit (89874, Thermo) were resuspended in 1x sample buffer (0.5 x TBE, 10% glycerol, 2% SDS) supplemented with protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (5 mM sodium fluoride, 1 mM sodium orthovanadate, 1 mM sodium pyrophosphate decahydrate, 1 mM β-glycerophophate) followed by protein concentration quantification using BCA method. Equal amounts of crude mitochondria were mixed with 1/10 volume gel loading dye (7025, NEB) and subjected to Semi-Denaturing Detergent Agarose Gel Electrophoresis (SDD-AGE). Samples were loaded onto a homemade vertical 1.5% agarose gel prepared using Novex empty gel cassettes (NC2015, Thermo), empty gel cassette combs (NC3515, Thermo) and Bio-Rad agarose (1613101, BioRad). After electrophoresis in the running buffer (1 x TBE and 0.1% SDS) for 40 min with a constant voltage of 100 V at 4°C, the proteins were transferred to 0.2 μM Amersham Protran Premium Nitrocellulose Western Blotting Membranes (10600004, GE) at 400 mA for 3 h in ice cold transfer buffer (10% methanol, 191 mM glycine, 25 mM tris-base, 0.1% SDS, pH 8.3) followed by immunoblotting.
Chen J., Harding S.M., Natesan R., Tian L., Benci J.L., Li W., Minn A.J., Asangani I.A, & Greenberg R.A. (2020). Cell Cycle Checkpoints Cooperate to Suppress DNA and RNA-Associated Molecular Pattern Recognition and Anti-Tumor Immune Responses. Cell reports, 32(9), 108080.
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2

In vivo MAVS Aggregation Assay

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
In vivo MAVS aggregation assay was performed according to a published protocol (Hou et al., 2011 (link)). Briefly, crude mitochondria isolated using mitochondria isolation kit (89874, Thermo) were resuspended in 1x sample buffer (0.5 x TBE, 10% glycerol, 2% SDS) supplemented with protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (5 mM sodium fluoride, 1 mM sodium orthovanadate, 1 mM sodium pyrophosphate decahydrate, 1 mM β-glycerophophate) followed by protein concentration quantification using BCA method. Equal amounts of crude mitochondria were mixed with 1/10 volume gel loading dye (7025, NEB) and subjected to Semi-Denaturing Detergent Agarose Gel Electrophoresis (SDD-AGE). Samples were loaded onto a homemade vertical 1.5% agarose gel prepared using Novex empty gel cassettes (NC2015, Thermo), empty gel cassette combs (NC3515, Thermo) and Bio-Rad agarose (1613101, BioRad). After electrophoresis in the running buffer (1 x TBE and 0.1% SDS) for 40 min with a constant voltage of 100 V at 4°C, the proteins were transferred to 0.2 μM Amersham Protran Premium Nitrocellulose Western Blotting Membranes (10600004, GE) at 400 mA for 3 h in ice cold transfer buffer (10% methanol, 191 mM glycine, 25 mM tris-base, 0.1% SDS, pH 8.3) followed by immunoblotting.
Chen J., Harding S.M., Natesan R., Tian L., Benci J.L., Li W., Minn A.J., Asangani I.A, & Greenberg R.A. (2020). Cell Cycle Checkpoints Cooperate to Suppress DNA and RNA-Associated Molecular Pattern Recognition and Anti-Tumor Immune Responses. Cell reports, 32(9), 108080.
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3

Western Blotting Protocol for Protein Analysis

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Western blotting was performed using standard methods. In brief, cells were collected by trypsinization, washed in PBS and lysed in NETN buffer (150 mM NaCl, 1% NP-40 alternative, 50 mM Tris pH 7.4) with turbo nuclease (Accelagen) in the presence of 1 mM MgCl2, protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (5 mM sodium fluoride, 1 mM sodium orthovanadate, 1 mM sodium pyrophosphate decahydrate, 1 mM β-glycerophophate) on ice for 30 min. Protein was quantified using Bradford method. Equal amount of protein was separated on 4%–12% Bis-Tris gels (Thermo Fisher Scientific) using MOPS/MES buffer and transferred to 0.2 μM Amersham Protran Premium Nitrocellulose Western Blotting Membranes (10600004, GE) at 350 mA for 2 h in ice cold transfer buffer (20% methanol, 191 mM glycine, 25 mM tris-base, 0.1% SDS, pH 8.3). Membrane were blocked in 3% non-fat milk in PBST (0.1% Tween 20) and incubated overnight at 4°C in primary antibody diluted with PBS containing 1% BSA. Membrane were washed in PBST for 20 min and incubated at room temperature for 2 h in secondary antibody (Amersham ECL HRP, GE) diluted with PBST containing 3% non-fat milk. Blots were developed using immobilon forte western HRP substrate (WBLUF0100, Millipore).
Chen J., Harding S.M., Natesan R., Tian L., Benci J.L., Li W., Minn A.J., Asangani I.A, & Greenberg R.A. (2020). Cell Cycle Checkpoints Cooperate to Suppress DNA and RNA-Associated Molecular Pattern Recognition and Anti-Tumor Immune Responses. Cell reports, 32(9), 108080.
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4

Western Blotting Protocol for Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blotting was performed using standard methods. In brief, cells were collected by trypsinization, washed in PBS and lysed in NETN buffer (150 mM NaCl, 1% NP-40 alternative, 50 mM Tris pH 7.4) with turbo nuclease (Accelagen) in the presence of 1 mM MgCl2, protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (5 mM sodium fluoride, 1 mM sodium orthovanadate, 1 mM sodium pyrophosphate decahydrate, 1 mM β-glycerophophate) on ice for 30 min. Protein was quantified using Bradford method. Equal amount of protein was separated on 4%–12% Bis-Tris gels (Thermo Fisher Scientific) using MOPS/MES buffer and transferred to 0.2 μM Amersham Protran Premium Nitrocellulose Western Blotting Membranes (10600004, GE) at 350 mA for 2 h in ice cold transfer buffer (20% methanol, 191 mM glycine, 25 mM tris-base, 0.1% SDS, pH 8.3). Membrane were blocked in 3% non-fat milk in PBST (0.1% Tween 20) and incubated overnight at 4°C in primary antibody diluted with PBS containing 1% BSA. Membrane were washed in PBST for 20 min and incubated at room temperature for 2 h in secondary antibody (Amersham ECL HRP, GE) diluted with PBST containing 3% non-fat milk. Blots were developed using immobilon forte western HRP substrate (WBLUF0100, Millipore).
Chen J., Harding S.M., Natesan R., Tian L., Benci J.L., Li W., Minn A.J., Asangani I.A, & Greenberg R.A. (2020). Cell Cycle Checkpoints Cooperate to Suppress DNA and RNA-Associated Molecular Pattern Recognition and Anti-Tumor Immune Responses. Cell reports, 32(9), 108080.
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