title: Optimization of protein buffer cocktails using Thermofluor.
abstract: The stability and homogeneity of a protein sample is strongly influenced by the composition of the buffer that the protein is in. A quick and easy approach to identify a buffer composition which increases the stability and possibly the conformational homogeneity of a protein sample is the fluorescence-based thermal-shift assay (Thermofluor). Here, a novel 96-condition screen for Thermofluor experiments is presented which consists of buffer and additive parts. The buffer screen comprises 23 different buffers and the additive screen includes small-molecule additives such as salts and nucleotide analogues. The utilization of small-molecule components which increase the thermal stability of a protein sample frequently results in a protein preparation of higher quality and quantity and ultimately also increases the chances of the protein crystallizing.
authors: EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, D-22603 Hamburg, Germany.
authors: Linda
authors: L
authors: Reinhard
authors: Hubert
authors: H
authors: Mayerhofer
authors: Arie
authors: A
authors: Geerlof
authors: Jochen
authors: J
authors: Mueller-Dieckmann
authors: Manfred S
authors: MS
authors: Weiss
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D002021:Buffers; D005453:Fluorescence; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D058767:Protein Unfolding; D011506:Proteins / Q000378:metabolism*; D012996:Solutions; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D002021:Buffers; D011506:Proteins; D012996:Solutions
keywords: Thermofluor; differential scanning fluorimetry; protein buffer cocktails; protein unfolding; small-molecule additives; thermal shift assay
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