Membrane:Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: JAK inhibitor withdrawal causes a transient pro-inflammatory cascade: A potential mechanism for major adverse cardiac events
Article Snippet: We loaded equal amounts of protein from lysates of SG-MSC or HUVECs to run on a tris- glycine-SDS 10% polyacrylamide gel (cat. #161-0374, Bio-Rad, Hercules, CA). .. This electrophoresis, in tris-glycine-SDS buffer at room temperature, was at 50V for ~45 minutes followed by ~135 minutes at ~90 V. Next, after incubating the gels on ice for 10 minutes, a ‘sandwich’ in a gel holder cassette – containing mesh, filter paper, the gel, the nitrocellulose membrane (0.2 μm), filter paper and mesh, from cathode to anode – was made for transfer of proteins from each gel to each membrane (cat. #1620112, BioRad, Hercules, CA) at 90 V for 90 minutes, with the chamber surrounded by ice in the cold room. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Secondary Structure Analysis of the C-Terminus of Gα-Interacting Vesicle Associated Protein Using Circular Dichroism Spectroscopy
Article Snippet: PVDF membrane, Immobilon-FL (EDM Millipore, Cat#IPFL00010) was activated with methanol then rinsed with transfer buffer. .. The gel was placed on the anode side while the membrane was on the cathode end, sandwiched between blot filter paper then foam pads stacked in the gel holder cassette (Bio-Rad, Catl703931/2/3). ..
Blocking Assay:Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Concentration Assay:Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Incubation:Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Gentle:Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Article Title: Methods of modulating seed and organ size in plants
Article Snippet: Air bubbles were removed from between the gel and membrane and then two further pieces of Whatman paper and a sponge were applied to the gel. .. This was enclosed in a gel holder cassette (from Bio-Rad Mini Trans-Blot® Cell kit), submerged in transfer buffer (25 mM TRIS, 192 mM glycine, 10% (v/v) methanol) and run at 90V for 70 minutes at 4° C. Following the transfer the membrane was washed for 10 minutes in 50 ml PBS (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, pH 7.3) at room temperature, before being agitated in 50 ml blocking solution (5% (w/v) milk powder, 0.1% (v/v) Tween-20) for either one hour at room temperature or overnight at 4° C. Primary antibodies were diluted to their appropriate concentration (see Table 2.9) in blocking solution and incubated with the membrane (10 ml per membrane with gentle agitation) for one hour before five washes with 50 ml PBST (140 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM KH2PO4, 0.1% (v/v) Tween-20, pH 7.3) at room temperature. ..
Electrophoresis:Article Title: JAK inhibitor withdrawal causes a transient pro-inflammatory cascade: A potential mechanism for major adverse cardiac events
Article Snippet: We loaded equal amounts of protein from lysates of SG-MSC or HUVECs to run on a tris- glycine-SDS 10% polyacrylamide gel (cat. #161-0374, Bio-Rad, Hercules, CA). .. This electrophoresis, in tris-glycine-SDS buffer at room temperature, was at 50V for ~45 minutes followed by ~135 minutes at ~90 V. Next, after incubating the gels on ice for 10 minutes, a ‘sandwich’ in a gel holder cassette – containing mesh, filter paper, the gel, the nitrocellulose membrane (0.2 μm), filter paper and mesh, from cathode to anode – was made for transfer of proteins from each gel to each membrane (cat. #1620112, BioRad, Hercules, CA) at 90 V for 90 minutes, with the chamber surrounded by ice in the cold room. ..
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