PureCube Compact Cartridge Ni-IDA
Order number: 30304
Description
Our PureCube Ni-IDA XL agarose resins are agarose resin beads with a diameter of ~400 µm. They are used for the purification of active His-tagged proteins from cells and extremely viscous cell media. Our year-long experience in manufacturing agarose resin lead to the high yield of 20 mg protein per ml resin, which is leading in the market compared to other Ni-IDA suppliers. PureCube Ni-IDA resins are suited for batch spin columns and FPLC. Their small diameter provides them with great mechanical stability.
For standard-sized beads with higher protein yield, we recommend our Ni-IDA agarose beads with 40 µm diameter.
Furthermore, you can also get the 40µm Ni-IDA beads pre-packed in a column/cartridge or as magnetic beads.
The PureCube Cartridge is competible with common FPLC systems like ÄKTA TM.
For standard-sized beads with higher protein yield, we recommend our Ni-IDA agarose beads with 40 µm diameter.
Furthermore, you can also get the 40µm Ni-IDA beads pre-packed in a column/cartridge or as magnetic beads.
The PureCube Cartridge is competible with common FPLC systems like ÄKTA TM.
Datasheets
Feature | |
---|---|
Usage | Specific binding and purification of 6x His tagged proteins |
Specificity | Affinity to His tagged proteins |
Binding capacity | >70 mg/mL |
Bead Ligand | Ni-IDA |
Bead size | 40 μm |
Filling quantity | Delivered as a 50 % suspension |
pH stability | 2-14 |
Other stabilities | 100% methanol, 100% ethanol, 8 M urea, 6 M guanidinium hydrochloride, 30% (v/v) acetonitrile |
Required equipment & materials |
|
Feature - Column | ||
---|---|---|
PureCube Compact Cartridge 1 mL | PureCube Compact Cartridge 5 mL | |
Bed Volume | 1 mL | 5 mL |
Max. Flow Rate | 6 mL/min | 6 mL/min |
Dimensions: diameter X length (mm) | 5 X 35 | 17 X 35 |
Body material | Polypropylene | Polypropylene |
Inlet | 10-32 UNF female thread | 10-32 UNF female thread |
Outlet | 10-32 UNF female thread | 10-32 UNF female thread |
Lab Results
Robust performance
Based on the same agarose matrix with high porosity and physical stability, PureCube Ni-IDA agarose exhibits a protein capacity of 50 mg protein per mL resin, which is competitive with that of products from market-leading providers. Figure 1 shows the SDS-PAGE analysis of 6-His chloramphenicol transferase (CAT) expressed in E. coli and purified on gravity flow columns filled with PureCUbe Ni-IDA Agarose or Competitor G Ni-Sepharose High-Performance resin. The amount of protein drawn down from the columns in 5 elution fractions was comparable for the two affinity resins (CL: cleared lysate; FT: flow-through; W1-2: wash fractions; E1-5: elution fractions). Table 1 compares additional parameters among PureCube Ni-IDA Agarose and two equivalent market-leading resins.
Based on the same agarose matrix with high porosity and physical stability, PureCube Ni-IDA agarose exhibits a protein capacity of 50 mg protein per mL resin, which is competitive with that of products from market-leading providers. Figure 1 shows the SDS-PAGE analysis of 6-His chloramphenicol transferase (CAT) expressed in E. coli and purified on gravity flow columns filled with PureCUbe Ni-IDA Agarose or Competitor G Ni-Sepharose High-Performance resin. The amount of protein drawn down from the columns in 5 elution fractions was comparable for the two affinity resins (CL: cleared lysate; FT: flow-through; W1-2: wash fractions; E1-5: elution fractions). Table 1 compares additional parameters among PureCube Ni-IDA Agarose and two equivalent market-leading resins.
Competitor | Particle Size | Metal ion capacity | Binding capacity | pH stability | Recommended flow rate | DTT stability | EDTA stability |
---|---|---|---|---|---|---|---|
Cube Biotech | 32-60 µm | >25 µmol/mL/mL | 70 mg/mL | 3.0-13.0 | 0.5-2.0mL/min (6.0 mL/min possible) |
10 mM | 1.0 mM |
Competitor G | average 90 µm | >30 µmol/mL | 15 mg/mL | 3.0-13.0 | 1.0 mL/min | No information | No information |
Competitor S | 45-160 µm | 6-18 µmol/mL | 70 mg/mL | Not provided | 1.0 mL/min | 5 mM | Not recommended |
Video
Video Guide - FPLC
FAQ
Can I get the datasheet for the Ni-IDA XL resin?
What are the reasons for non-specific binding?
There are some protein that can bind to Ni-IDA even without having a His-tag. But to a lesser extent. Washing with NaOH after elution of your protein of interest removes unspecific bound proteins from your resin.
I want to use a high concentration of EDTA and DTT. Is it possible to use Ni-IDA from Cube Biotech?
No, it is not recommended because nickel-ions are reduced with DTT or dissolved with EDTA. If you want to use high concentrations of EDTA and DTT you should use our Indigo resin.
How is the capacity at high flow rates?
If higher flow rates are desired we recommend using beads with bigger diameters. We offer Ni-IDA beads with mean diameters of 40µm, and 400µm (XL).
With each size increase, the flow rates also increase due to the proportionally increasing space between the beads. However, the surface of the beads does not increase at the same speed as the diameter (square-cube-law). That results in decreasing amounts of purified protein per mL beads while increasing the bead sizes.
For 40µm we have average purification amounts of ~70 µg protein/mL beads. With 400 µm (XL) beads, this decreases to 20 µg/mL beads.
We recommend reading the corresponding section of the "Introduction to agarose matrixes" guide on this subject for more detailed information.
With each size increase, the flow rates also increase due to the proportionally increasing space between the beads. However, the surface of the beads does not increase at the same speed as the diameter (square-cube-law). That results in decreasing amounts of purified protein per mL beads while increasing the bead sizes.
For 40µm we have average purification amounts of ~70 µg protein/mL beads. With 400 µm (XL) beads, this decreases to 20 µg/mL beads.
We recommend reading the corresponding section of the "Introduction to agarose matrixes" guide on this subject for more detailed information.
After using DTT my resin changed color. How to regenerate it?
The DTT has probably destroyed your beads. Ni-IDA beads only have a very limited DTT tolerance. However, you can regenerate them to regain their functionality. Please read our detailed protocol for more information regarding this. It is linked above.
However, we would recommend using Ni-INDIGO products instead. They work with the same buffers and protocols as the Ni-NTA products but have a DTT tolerance of 20 mM.
However, we would recommend using Ni-INDIGO products instead. They work with the same buffers and protocols as the Ni-NTA products but have a DTT tolerance of 20 mM.