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High-throughput biotinylation of proteins

, : High-throughput biotinylation of proteins. Methods in Molecular Biology 498: 185-196

One of the more useful tags for a protein in biochemical experiments is biotin, because of its femtomolar dissociation constant with streptavidin or avidin. Robust methodologies have been developed for other the in vivo addition of a single biotin to recombinant protein or the in vitro enzymatic or chemical addition of biotin to a protein. Such modified proteins can be used in a variety of experiments, such as affinity selection of phage-displayed peptides or antibodies, pull-down of interacting proteins from cell lysates, or displaying proteins on arrays. We present three complementary approaches for biotinylating proteins in vivo in Escherichia coli or in vitro using chemical or enzymatical reactions all of which can be scaled up to tag large numbers of proteins in parallel.


PMID: 18988027

DOI: 10.1007/978-1-59745-196-3_13

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Edel' K.E.; Gorelov A.N.; Gladkov M.A., 1987: Milk productivity of cows as a function of the age and milk productivity of their mothers. It is found in the investigations conducted with 400 cows in the State Breeding Farm "Zarja Kommunisma" (Domodedovsky district, Moscow region) that the effect of dams' age on the milk yield of their daughters in a high yielding herd...

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Prieto, S.; Bouillaud, F.; Ricquier, D.; Rial, E., 1992: Activation by ATP of a proton-conducting pathway in yeast mitochondria. The growth of Saccharomyces cerevisiae cells under aerobic conditions, in the presence of an energy-rich source, leads to production of an excess of NAD(P)H. Since the redox balance must be maintained, it has been postulated that NAD(P)H reoxidati...

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Heintz, B.; Stöcker, G.; Mrowka, C.; Rentz, U.; Melzer, H.; Stickeler, E.; Sieberth, H.G.; Greiling, H.; Haubeck, H.D., 1995: Decreased glomerular basement membrane heparan sulfate proteoglycan in essential hypertension. Heparan sulfate proteoglycans are major components of the glomerular basement membrane and play a key role in the molecular organization and function of the basement membrane. Moreover, their presence is essential for maintenance of the selective...

Anonymous, 1981: Symposium on orthodontics. Dental Clinics of North America 25(1): 1-205

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Mckinney, H.H.; Sando, W.J., 1951: Susceptibility and resistance to the wheat streak-mosaic virus in the genera Triticum, Agropyron, Secale, and certain hybrids. All the selections that have shown high mosaic resistance or possible immunity will require many additional crosses with wheat before desirable commercial wheats can be obtained.