The Thermofluor Screens JBScreen FUNDAMENT and JBScreen SPECIFIC allow identification of protein-stabilizing buffer conditions which is pivotal for protein purification, characterization and crystallization.
Undesired overlay of screening of interdependent variables is prevented by strictly categorizing stability screening of proteins into

1. FUNDAMENTAL factors that influence the whole protein molecule(pH and ionic strength)
2. SPECIFIC factors that affect energetically important hot spots on the protein (substrates and their analogs, cations, anions, …)

The protein’s melting temperature (Tm) is used as reporter for protein stability and determined by a thermal shift assay (96-well plate format) monitoring the unfolding of a protein in a temperature-dependent manner. The higher the Tm, the higher is the thermostability of the protein in that specific environment.

JBScreen Thermofluor FUNDAMENT and SPECIFIC are provided in a deep-well block at 0.5 ml each, which allows a flexible application of this alternative approach.

Example: Stabilizing effects on α-Chymotrypsinogen A are directly correlated with crystallizability

α-Chymotrypsinogen A was applied in JBScreen Thermofluor FUNDAMENT and SPECIFIC.
Stabilizing conditions (high ionic strength, neutral pH, CaCl₂ & FeCl₃) were combined without further optimization. A crystallization screen was set up:

• without any specific ion
• with 20 mM CaCl₂
• with 10 mM FeCl₃

类别：结晶筛
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Thermofluor Screens JBScreen FUNDAMENT 和 JBScreen SPECIFIC 允许鉴定蛋白质稳定缓冲液条件，这对于蛋白质纯化、表征和结晶至关重要。
通过将蛋白质的稳定性筛选严格分类为

1. 影响整个蛋白质分子的基本因素（pH 值和离子强度）
2. 影响蛋白质能量上重要热点的特定因素（底物及其类似物、阳离子、阴离子……）

蛋白质的解链温度 (Tm) 用作蛋白质稳定性的报告基因，并通过热位移测定（96 孔板格式）以温度依赖性方式监测蛋白质的解折叠来确定。 Tm 越高，蛋白质在特定环境中的热稳定性就越高。

JBScreen Thermofluor FUNDAMENT 和 SPECIFIC 以每个 0.5 ml 的深孔块提供，允许灵活应用这种替代方法。

示例：对 α-胰凝乳蛋白酶原 A 的稳定作用与结晶性直接相关

α-糜蛋白酶原 A 应用于 JBScreen Thermofluor FUNDAMENT 和 SPECIFIC。
稳定条件（高离子强度、中性 pH 值、CaCl2 和 FeCl3）组合在一起，无需进一步优化。设置结晶筛：

没有任何特定的离子
含 20 mM CaCl2
含 10 mM FeCl3