Lb media

Manufactured by MP Biomedicals

Sourced in United States

LB media is a commonly used microbiology growth medium designed to support the cultivation of a wide range of bacteria. It provides essential nutrients and growth factors necessary for the proliferation of bacterial cultures.

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Lab products found in correlation

Histrap ff, by GE Healthcare (1 mentions) Superdex 75 column, by GE Healthcare (1 mentions) Ni nta bead, by Qiagen (1 mentions) Protease inhibitor, by Roche (1 mentions) Gblock, by Integrated DNA Technologies (1 mentions) Gc rich pcr system, by Roche (1 mentions) 2 propanol, by Thermo Fisher Scientific (1 mentions) Lc ms ultra grade formic acid fluka, by Merck Group (1 mentions) Ultrafree mc gv centrifugal filters, by Merck Group (1 mentions)

2 protocols using lb media

Protein Purification and Characterization

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Genes were obtained from GenScript directly in pET21b or synthesized as gBlocks (Integrated DNA Technologies) and cloned into pET29b. For the clones used for folding assessments, plasmids were transformed into E. coli BL21(DE3) strain (Novagen, cat. no. 69450) for IPTG induced expression in LB media (MP Biomedicals, cat. no. 113002056) at 18 °C overnight. Cell pellets were collected by centrifugation at 6000 g for 20 minutes, resuspended in Tris buffer (20 mM Tris pH 8, 150 mM NaCl) with protease inhibitors (Roche, cat. no. 11697498001) and lysed by sonication. Ni-NTA beads (Qiagen, cat. no. 30410) or HisTrap FF (GE Life Sciences, cat. no. 17-5255-01) were used to purify the proteins, with imidazole elution concentrations between 10 mM to 500 mM. Size exclusion by gel filtration steps were performed on ÄKTAxpress units (GE Life Sciences) using Superdex 75 columns (GE Life Science, cat. no. 17-5174-01) with 50 mM sodium phosphate buffer pH 8 with 150 mM NaCl.

Huang P.S., Feldmeier K., Parmeggiani F., Velasco D.A., Höcker B, & Baker D. (2015). De novo design of a four-fold symmetric TIM-barrel protein with atomic-level accuracy. Nature chemical biology, 12(1), 29-34.

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Proteomic Profiling of Recombinant Proteins

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J.T. Baker Ultra LC/MS-grade acetonitrile (ACN) and water were purchased from VWR International (Radnor, PA). LC/MS Ultra-grade formic acid (Fluka) was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Roswell Park.
Memorial Institute (RPMI) media (Gibco), pBAD Directional TOPO® Expression Kit, and ProBond™ Purification System were purchased from Invitrogcn (Carlsbad. CA, USA). DL- dithiothreitol (DTT), MgS0₄, and ¹³C₆, ¹⁵N₄-L-arginine hydrochloride were purchased from Sigma-Aldrich (St. Louis, MO, USA). LB media (MP Biomedicals), GC-Rich PCR system (Roche), 4-(2-hydroxyethyl)-l -pipcrazineethancsulfonic acid (HEPES) at pH 8.4, 2-propanol (Fisher Chemical), and borate buffer at pH 9.5 (R1CCA Chemical) were purchased from Fisher Scientific (Pittsburgh, PA, USA). The Quick Blunting kit and restriction enzymes (EcoRI, EcoRV, HindIII, SphI) were purchased from New England Biosciences (Ipswich, MA, USA). 4-Fluoro-7-nitro-2,l,3-bcnzoxadiazole (NBDF) was purchased from Dojindo Laboratories (Rockville, MD, USA). Amicon Ultra 3 kDa MW cutoff centrifugal filters and Ultrafree-MC GV centrifugal filters were purchased from Millipore (Milford, MA, USA). Polypropylene auto-sampler tubes with PTFE/silicone septa were purchased from Waters (Milford, MA, USA).

Dalluge J.J., McCurtain J.L., Gilbertsen A.J., Kalstabakken K.A, & Williams B.J. (2015). Determination of agmatine using isotope dilution UPLC-tandem mass spectrometry: application to the characterization of the arginine decarboxylase pathway in Pseudomonas aeruginosa. Analytical and bioanalytical chemistry, 407(18), 5513-5519.

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