His-纯化试剂盒.pdfVIP

GE Healthcare Instructions 28-4010-39 AC His Buffer Kit Intended use His Buffer Kit is intended for research use only, and should not be used in any clinical or in vitro procedures for diagnostic purposes. His Buffer Kit contains • Phosphate buffer 8 x stock solution, pH 7.4 (160 mM sodium phosphate, 4 M NaCl). Volume 2 x 100 ml. • 2 M imidazole, pH 7.4. Volume 1 x 100 ml. • Instructions for use. Purpose His Buffer Kit contains stock solutions of binding and elution buffers for purification of histidine-tagged proteins. The kit eliminates time-consuming buffer preparation and thus promotes fast, reproducible and convenient purification work. His Buffer Kit can be used in combination with a number of column types, such as His SpinTrap™, His MultiTrap™ FF, His MultiTrap HP, His GraviTrap™, HisTrap™ FF, HisTrap FF crude and HisTrap HP. Table of contents 1. Principle 2 2. Buffer preparation 3 3. Optimization of imidazole concentration 5 4. Purification protocol 6 5. Ordering information 6 1 Principle Imidazole is commonly used to elute histidine-tagged proteins from IMAC (immobilized metal affinity chromatography) media. To minimize the binding of unwanted host cell proteins, imidazole is also used at a low concentration in the binding buffer. For this reason, it is important to include imidazole in the sample at the same concentration as in the binding buffer. 20 mM concentration of imidazole in the binding buffer commonly gives good purification results but the purification is protein-dependent. Optimization of the imidazole concentration may be required, see Section 3. 2 28-4010-39 AC 2 Buffer preparation The choice of optimal imid