Immunoblot analysis was performed as described previously114 (link) with minor changes. Samples were separated on a Mini-PROTEAN TGX 5%–20% Tris-Glycine gel (Bio-Rad) and transferred to a nitrocellulose membrane (Thermo Fisher Scientific). Membranes were blocked in 1X TBST containing 5% milk, probed with a 1:2,000 dilution of polyclonal α-PotA65 followed by a 1:10,000 dilution of peroxidase labeled α-mouse (GE Healthcare) and developed with SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) on a Bio-Rad ChemiDoc MP Imaging System.
Mini protean tgx 5 20 tris glycine gel
Manufactured by Bio-Rad
The Mini-PROTEAN TGX 5%–20% Tris-Glycine gel is a precast polyacrylamide gel used for protein separation and analysis. It features a gradient of 5% to 20% polyacrylamide concentration, which allows for the separation of a wide range of protein molecular weights. The gel is designed for use with the Mini-PROTEAN electrophoresis system.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by Thermo Fisher Scientific (5 mentions)
Chemidoc mp imaging system, by Bio-Rad (3 mentions)
Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (3 mentions)
Peroxidase labeled α mouse, by GE Healthcare (2 mentions)
Amersham ecl western blotting detection kit, by GE Healthcare (2 mentions)
Amersham ecl western blotting, by Cytiva (2 mentions)
Laemmli buffer, by Bio-Rad (1 mentions)
Non fat milk, by Bio-Rad (1 mentions)
5 protocols using mini protean tgx 5 20 tris glycine gel
1
Immunoblot Analysis of PotA Protein
2
Immunoblot Analysis of PotA Protein
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Immunoblot analysis was performed as described previously with minor changes (Zhang et al., 2002 (link)). Samples were separated on a Mini-PROTEAN TGX 5%–20% Tris-Glycine gel (Bio-Rad) and transferred to a nitrocellulose membrane (Thermo Fisher Scientific). Membranes were blocked in 1X TBST containing 5% milk, probed with a 1:2,000 dilution of polyclonal α-PotA (Lin et al., 2017 ) followed by a 1:10000 dilution of peroxidase labeled α-mouse (GE Healthcare) and developed with SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) on a Bio-Rad ChemiDoc MP Imaging System.
3
Immunoblot Analysis of Protein Samples
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Immunoblot analysis was performed as described previously with minor changes (Zhang et al., 2002 (link)). Samples were separated on a Mini-PROTEAN TGX 5–20% Tris-Glycine gel (Bio-Rad) and transferred to a nitrocellulose membrane (Thermo Fisher Scientific). Membranes were blocked in 1X TBST containing 5% milk, probed with a 1:2000 dilution of monoclonal α-FLAG-HRP (Sigma) or a 1:500 dilution of polyclonal α-OmpC follwed by a 1:1000 dilution of peroxidase labeled α-rabbit and developed with SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific) on a Bio-Rad ChemiDoc MP Imaging System.
4
Immunoblot Analysis of Hfq and ProQ
5
Western Blot Analysis of Bacterial Proteins
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The cell pellet from 1 ml of cells grown in the indicated medium was resuspended in 1X PBS (KD Medical) , 7 µl of 2X Laemmli buffer (BioRad) and 2 µl of b-mercaptoethanol, and 10 µl were loaded on a Mini-PROTEAN TGX 5%-20% Tris-Glycine gel (Bio-Rad) and run in 1X Tris Glycine-SDS (KD Medical) buffer. The proteins were electro-transferred to nitrocellulose membranes (Invitrogen) for 1 h at 100 V. Membranes were blocked with 5% non-fat milk (BioRad) in 1X PBS with 0.1% of Tween 20 (PBS-T) for 1 h and probed with a 1:3,000 dilution of a-FLAG-HRP antiserum (Sigma), 1:1,000 dilution of a-AzuC antiserum (New England Peptide); 1:1,000 dilution of a-His-HRP antiserum (Qiagen), or 1:1,000 dilution of a-OmpA antiserum (Antibody Research Corporation) in the same PBS-T buffer with 5% milk for 1 h.
After the incubation with the a-AzuC and a-OmpA antiserum, membranes were incubated with a 1:2,000 dilution of HRP-labelled anti-rabbit antibody (Life Technologies). All blots were washed 4X with PBS-T and then developed with a Amersham ECL Western Blotting Detection Kit (GE Healthcare).
After the incubation with the a-AzuC and a-OmpA antiserum, membranes were incubated with a 1:2,000 dilution of HRP-labelled anti-rabbit antibody (Life Technologies). All blots were washed 4X with PBS-T and then developed with a Amersham ECL Western Blotting Detection Kit (GE Healthcare).
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