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Lowry Protein Assay Lowry protein esej

Protein Determination by the Lowry Assay Određivanje proteina po Lowry esej

The Lowry protein assay method for protein concentration determination is one of the most venerable and widely-used protein assays. The Lowry protein esej metoda za određivanje koncentracije proteina jedan je od najatraktivnijih časni i naširoko koristi protein-eseji. The Lowry method was first described in 1951 by Lowry et al. The Lowry metoda je prvi put opisana u 1951 by Lowry et al. (Lowry et al., 1951). (Lowry et al., 1951).

Hydrolysis is probably the most accurate method of determining protein concentration  followed by amino acid analysis. Hydrolysis je vjerojatno najpouzdanija metoda određivanja koncentracije proteina slijedi analiza aminokiselina. Most other methods are sensitive to the Većina druge metode su osjetljive na
amino acid composition of the protein, and absolute concentrations cannot be obtained. aminokiselina sastav proteina, a apsolutna koncentracija ne može biti dobiven. The Lowry procedure is sensitive, and is moderately constant from protein to protein. The Lowry postupak je osjetljivo, te je umjereno konstanta iz bjelančevina u proteinu. The Lowry protein estimation  has been so widely used that it is a completely acceptable alternative to a rigorous absolute determination in almost all circumstances in which protein mixtures or crude extracts are involved. The Lowry protein procjena je tako široko koristi da je potpuno prihvatljivo alternativno na strog apsolutnu odlučnost u gotovo svim okolnostima u kojima protein mješavina ili sirova ekstrakti su uključeni.

The method is based on both the Biuret reaction, in which the peptide bonds of proteins react with copper under alkaline conditions to produce Cu+, which reacts with Metoda se temelji na oba Biuret reakciji, u kojoj su obveznice peptida proteina reagira s bakrom alkalni pod uvjetima za proizvodnju Cu +, koja reagira s
the Folin reagent, and the Folin–Ciocalteau reaction, which is poorly understood but in essence phosphomolybdotungstate is reduced to heteropolymolybdenum blue by the copper-catalyzed oxidation of aromatic amino acids. Folin, reagens, i Folin-Ciocalteau reakcija, koja je slabo razumio, ali u sustini phosphomolybdotungstate je sveden na heteropolymolybdenum plava od bakra-oxidation catalyzed aromatičnih aminokiseline. The reactions result in a strong blue color, which depends partly on the tyrosine and tryptophan content. Reakcije rezultat u jake plave boje, koja dijelom ovisi o tirozinska i triptofan sadržaj. The method is sensitive down to about 0.01 mg of protein/mL, and is best used on solutions with concentrations in the range 0.01–1.0 mg/mL of protein. Metoda je osjetljiva dolje na oko 0,01 mg od proteina / mL, te je najbolje koristiti na rješenja s koncentracijama u rasponu 0.01-1.0 mg / mL of protein.

Chemicals and Substances that Interfere with the Lowry Assay Kemikalije i supstance koje interferirati s Lowry esej

The Lowry method for protein concentration determination is widely used, however can be subject to interference by many substances and chemicals. The Lowry metoda za određivanje koncentracije proteina je naširoko koristi, ali može biti predmetom interference by mnoge tvari i kemikalija.

Chemicals that are known to interefere with the Lowry Assay: Kemikalije koje se zna da interefere s Lowry esej:

• Barbital
• CAPS Kape
• Cesium Chloride Cezij Chloride
• Citrate
• Cysteine
• Diethanolamine
• Dithiothreitol
• EDTA
• EGTA
• HEPES
• mercaptoethanol
• Nonidet P-40 Nonidet P-40
• Phenol
• Polyvinyl pyrrolidone Polivinil pyrrolidone
• Sodium Deoxycholate Natrij Deoxycholate
• Sodium Salicylate Natrij Salicylate
• Thimerosol
• Tricine
• TRIS
• Triton X-100 Triton X-100

Lowry Method References Lowry Metoda Reference

Lowry et al. J. Lowry et al. J. Biol. Biol. Chem . Chem. 193: 265-275 (1951) 193: 265-275 (1951)

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