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HisPur™ Cobalt Purification Kit, 3 mL - FAQs

查看更多产品信息 HisPur™ Cobalt Purification Kit, 3 mL - FAQs (90092)

4 个常见问题解答

细菌无法用B-PER试剂裂解，这是什么原因所致？

在富营养培养基（高糖）中生长的细菌往往难以裂解。您可以尝试减少葡萄糖浓度。B-PER试剂能够裂解包括E. coli，Acetinobacter，Archaebacteria，S. aureus，H. pylori，昆虫杆状病毒，线虫和昆虫细胞在内的细菌和细胞。B-PER试剂可以裂解一些革兰氏阳性菌，但是裂解革兰氏阴性细菌的效率通常更高。如果对某一特定细菌菌株裂解效果不佳，则请在裂解前冷冻细菌。添加溶菌酶是最有效的提取手段，但是对于一些过表达蛋白则不需要加入溶菌酶。

The bacteria do not lyse with B-PER reagent. What is the problem?

Bacteria grown in rich medium (high glucose) are difficult to lyse. Try reducing the glucose concentration. Bacteria and organisms lysed with B-PER Reagent include E. coli, Acetinobacter, Archaebacteria, S. aureus, H. pylori, Baculovirus, nematodes, and insect cells. B-PER Reagent can lyse some gram-positive bacteria, but tends to be more efficient with gram-negative bacteria. If lysis is inefficient for a particular bacterial strain, freeze cells before extraction. Add lysozyme for the most efficient extraction, however, for some over-expressed proteins, the addition of lysozyme is not required.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What is the difference between HisPur Nickel Resin and HisPur Cobalt Resin?

Nickel has a higher affinity for histidines than does cobalt. It binds multiple histidines more tightly than cobalt, and requires more stringent conditions than is necessary for cobalt. The lower affinity of cobalt for multiple histidines typically results in less nonspecific binding of histidine-rich proteins that lack a his-tag compared to nickel resins.

HisPur Ni-NTA resin has a high capacity of up to 60 mg of 6xHis-tagged protein per milliliter. It is a versatile resin that can work under both native and denaturing conditions, and can also be used with a variety of lysis reagents and buffer additives.

HisPur Cobalt resin utilizes proprietary tetradentate chelating resin charged with cobalt. This system recovers highly purified protein with lower imidazole concentrations and has low metal leeching properties.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Is the rapid equilibrium dialysis (RED) system compatible with automation?

Yes. The Teflon Base Plate has the standard 96-well footprint. It can be used with any system compatible with standardized 96-well ELISA plates with 9 by 9 mm well spacing. The single-use format for the RED Device is especially convenient for labs using radioactive materials because the plate can easily be disposed of to minimize contamination and the need for cleaning.