PureCube Compact Cartridge Ni-IDA

Order number: 30304

€201.00*

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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.
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
  • FPLC hardware like e.g. AEKTA
  • Binding Buffer
  • Elution Buffer


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.
PureCube Ni-IDA Performance compared to competitors
Figure 1: The binding capacity of PureCube Ni-IDA Agarose is comparable to resins from a leading supplier. SDS-PAGE of CAT purified in gravity columns with PureCube Ni-IDA Agarose and Ni resin from Competitor G show elution fractions (E1-E5) with similar protein yields.
Table 1: Comparison of three Ni-IDA manufacturers on the market.
Competitor Particle SizeMetal ion capacityBinding capacitypH stabilityRecommended flow rateDTT stabilityEDTA 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.

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.