All chemicals used were purchased from Sigma (St. Louis, MO) unless otherwise noted. Nitrocefin was purchased from TOKU-E (Bellingham, WA). Bovine Serum Albumin (BSA) was purchased from New England Biolabs (Ipswich, MA). BL21 competent cells purchased from Agilent Technologies (Santa Clara, CA). pDIMC8-RG13-AND2, -YES, and -ORN2, plasmids that contain the RG13 disulfide variant genes, were previously described (Choi and Ostermeier, 2014 (link)). Electrochemical cell kit was purchased from BASi (West Lafayette, IN). The potentiometer was from purchased from Princeton Applied Research (Oak Ridge, TN).
Bl21 competent cells
Manufactured by Agilent Technologies
BL21 competent cells are a strain of E. coli bacteria commonly used in molecular biology and protein expression experiments. They are designed to facilitate the efficient production of recombinant proteins.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocefin, by Merck Group (2 mentions)
Quikchange mutagenesis procedure, by Agilent Technologies (2 mentions)
Bovine serum albumin (bsa), by New England Biolabs (1 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
Gst agarose beads, by Thermo Fisher Scientific (1 mentions)
Isopropyl β d thiogalactoside, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
5 protocols using bl21 competent cells
1
Biosensor-based Antibiotic Susceptibility Assay
2
Recombinant Sox2 Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial expression constructs (PGEX-6P-1-Sox2) containing the indicated genes were transformed into BL21-competent cells (Agilent Technologies). Cells were induced to overexpress protein under 0.1 mM isopropyl-b-D-thiogalactoside (Sigma-Aldrich) at 16 °C for 16-18 h. Cells were resuspended in PBS containing Protease Inhibitor Cocktail (Roche) and Phosphatase Inhibitor Cocktail (Roche), followed by ultrasonication. The proteins were purified using GST agarose beads according to the manufacturer's protocol (Thermo Fisher). Three replicates were performed.
3
Mutagenesis and Peptide Synthesis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals used were purchased from Sigma (St. Louis, MO) unless otherwise noted. NMR tubes were purchased from Shigemi. (Allison Park, PA). Nitrocefin was purchased from EMD Millipore (Billerica, MA). BL21 competent cells purchased from Agilent Technologies. Plasmids pDIMC8-c4h and pDIMC8-MBP that contain the c4 and malE genes respectively were previously described9 (link),10 (link). Plasmid pDIMC8-c4h was used as a template plasmid DNA to make linker variants using the PCR-based QuikChange mutagenesis procedure (Agilent Technologies). DNA sequencing of 2013 to 2061 bp regions containing the targeted codon confirmed mutant clones (Genewiz). The WA-(EAQA)7 peptide (sequence: WAEAQAEAQAEAQAEAQAEAQAEAQAEAQA) was custom synthesized with 98% purity by Peptide 2.0. synthesis, and peptide ends were modified by N-terminal acetylation and C-terminal amidation.
4
Bacterial Protein Expression and Purification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial expression constructs (pGEX-4T-2) containing the indicated genes were transformed into BL21-competent cells (Agilent Technologies). Cells were induced to protein overexpression under 0.1–0.5 mM isopropyl-b-D-thiogalactoside at 37 °C. Cells were resuspended in PBS containing 0.5% Triton X-100, 5 mM β-mercaptoethanol, 2 mM EDTA and 1 mM PMSF, followed by ultrasonication. The proteins were purified by a single step using glutathione bead according to the manufacturer's protocol (GE Health Science).
5
Mutagenesis and Peptide Synthesis Protocol
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!