HisPur™ Ni-NTA Resin, 10 mL - FAQs

View additional product information for HisPur™ Ni-NTA Resin - FAQs (88223, 88222, 88221)

6 product FAQs found

Are there other sequences that can bind to nickel more tightly than 6xHis-tagged proteins and how can they be eluted.

Yes, 7xHis-tagged proteins, proteins naturally high in histidine, and other combinations of His and other amino acids will bind. To elute them, you have to increase the concentration of imidazole. Generally these peptides will not contaminate your fraction since they remain on the column. However after multiple uses of the same column, these peptides may reduce the binding capacity of the column.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can the HisPur Ni-NTA Spin plates (Cat. No. 88230) be regenerated and reused, or are they for single-use only?

The resin can be re-used at least five times; this is also for the resin in the plates. Regeneration is possible using this procedure (https://assets.thermofisher.com/TFS-Assets%2FLSG%2Fmanuals%2FMAN0011700_HisPur_NiNTA_Resin_UG.pdf) Since the resin bed volume in each well is 50 µL, you can use 250 µL twice to give a total of 10 resin-bed volume.

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

How do I perform Ni-NTA bead regeneration with the HisPur Ni-NTA Resin (Cat. No. 88221)?

Please refer to page 2 (buffer preparation) and page 3 (procedure) of the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0011700_HisPur_NiNTA_Resin_UG.pdf) for information regarding bead regeneration.

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

Can ProBond or Ni-NTA beads be used for large-scale preparations?

ProBond and Ni-NTA beads can be used in FPLC columns. However, the beads can only withstand low pressure (~43.5 psi max).

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

What should the typical protein recovery be when using the Probond Purification System or Ni-NTA Purification system?

Both systems are qualified by purifying 2 mg of myoglobin protein on a column and performing a Bradford assay. Protein recovery must be 75% or higher.

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.