"The combination of high protein binding capacity and ease of use makes the PureCube Ni-INDIGO resin our go-to product when it comes to Ni-affinity purification.
The resin seems to retain its full protein binding capacity even after 30 uses when each purification is followed by the recommended CIP protocol. The INDIGO technology allows you to clean the resin with 0.5 M NaOH without stripping the Nickel thus eliminating tedious resin metal ion stripping and recharging. We would highly recommend PureCube Ni-INDIGO resin to anyone doing Ni-Affinity protein purification."Time-saving PureCube Purification Technologies
Benefit from affinity beads developed by experts with 30 years of experience
Recombinant protein production | Membrane protein purification | Protein-protein interaction studies | Co-immunoprecipitation | Mass spectrometry sample preparation | ELISA | Western Blot detection | Protein localization |
Protein research is driven by our shared passion for discovery and methodological improvement. During protein purification, our customers often report challenges such as
high levels of unspecific binding, aggregated protein, nickel-ion leakage, the need for specific customized resin, low yield, and purity. These challenges drive us to develop innovative products that transcend conventional protein purification methods, enabling customers to make valuable contributions to our field.
Technology 1: His-tag affinity
Highest protein yields due to Ni-Leakage resistant beads
PureCube Ni-INDIGO Agarose and MagBeads
Figure 1: Yield of recovered GFP in a purification batch process with PureCube Ni-INDIGO Agarose with different tested additives 0.1M Citrate, 20 mM DTT, 6 mM GuHCl, 8 M GuHCl, 10 TCEP, 2% Titron, 8 M Urea, 20 mM EDTA.
Figure 2: Comparison of the protein purity of a His-tag purification assay. PureCube Ni-INDIGO agarose from Cube Biotech in contrast to Ni-NTA agarose from a market competitor "T". Both purifications were performed under the same conditions.
Figure 3: Protein yield of His-tagged protein compared between PureCube Ni-INDIGO based beads and competitor products at 20 mM EDTA and 10 mM DTT.
PureCube Ni-INDIGO-Technology in research
Michael Crone, PhD,
Solena Materials,
London
"We tested three types of (...) IMAC resins: Ni-NTA Agarose (Qiagen), Ni Sepharose excel (Cytiva), INDIGO-Ni agarose resin. We found that the cell media leached the nickel ions from the Ni-NTA Agarose resin, significantly reducing the final protein yield.
Excel and INDIGO-Ni are both chemical resistant resins, and both showed no observable metal ion leaching. We decided to proceed with our experiments using the INDIGO-Ni resin, as the Excel resin co-purified more impurities (...)"
Stuble et al., 2022
Excel and INDIGO-Ni are both chemical resistant resins, and both showed no observable metal ion leaching. We decided to proceed with our experiments using the INDIGO-Ni resin, as the Excel resin co-purified more impurities (...)"
Stuble et al., 2022
PureCube Ni-INDIGO was utilized to obtain high-purity Mpro, which is necessary for testing the efficacy of the newly designed inhibitors. "High purity protein was eluted in a buffer of 50 mM Tris (pH 8), 250 mM Imidazole, and 300 mM NaCl. Eluted fractions with high protein content were pooled and buffer exchanged into HRV protease cleavage buffer..."
Westberg, et al. 2020
Westberg, et al. 2020
Technology 2: His-tag affinity
Evolution of Ni-NTA: Beads with the highest ligand density
PureCube Ni-NTA Agarose and MagBeads
Figure 4: His-tagged GFP was purified from 1. E.coli cell lysates, and 2. Insect cell lysates on His affinity resins in batch format. PureCube Ni-NTA Agarose was compared to conventional Ni-NTA resins of leading competitors, we refer to them as “A“, “B“, and “C“. Recovered GFP was quantified by absorption at 488 nm. Binding E.coli lysates: 50 mM NaH2PO4, 300 mM NaCl, 10 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4; Binding Insect cell supernatants: Sample as is with 10 mM imidazole added; Wash: 50 mM NaH2PO4, 300 mM NaCl, 20 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4; Elution: 50 mM NaH2PO4, 300 mM NaCl, 250 mM imidazole, 1 mM EDTA, 10 mM DTT, pH 7.4
Figure 5: Over 20% more yield was obtained with PureCube Ni-NTA Agarose. SDS-PAGE of GFP expressed in E. coli and purified in gravity columns with PureCube Ni-NTA Agarose and Ni-NTA resin from Competitor G, and Competitor Q. 80 mg/mL protein yield was obtained with PureCube Ni-NTA Agarose (E1–E4, Cube) compared to 65 and 48 mg/mL, respectively, with the widely used alternative resins G and Q (E1–E4, Competitor G / Competitor Q).
Technology 3: Rho1D4-tag Affinity
Unmatched purity for low-abundance proteins
HighSpec Rho1D4 Agarose
Fig. 6: Purification of chemokine receptor 4 (CXCR4) using PureCube Rho1D4 Agarose. Total lysate (TL) was solubilized with Fos-Choline-14, and the soluble fraction (SF) was incubated on an immunoaffinity column loaded with rho1D4 antibody. The concentration of eluted CXCR4 ranged 0.6-1.0 mg/mL as determined spectrophotometrically.
"My working group uses rho1D4 resins
from Cube Biotech to purify membrane
proteins. For our applications we need very high purity. The rho1D4-tag provides
high specificity and yield at the same time. (...)
We will certainly use it again in the future! "
Prof. Dr. Jörg Labahn, Group leader at the Centre for Structural Systems Biology
Technology 4: Strep-tag® Affinity
The best balance of specificity and purity
PureCube HiCap StrepTactin® Agarose and Magbeads
Figure 7: Binding capacity during purification of CD45 Nanobody Twin-Strep-tag® 16.2 kDa and CD56 Antigen Twin-Strep-tag® 82.6 kDa in gravity flow
Figure 8: Purification of GFP-Twin-Strep-tag® from E.coli lysate using HiCap Strep-Tactin®
Technology 5: Active surface groups
Unimagined possibilities with customized beads
Activated Resins & MagBeads
- Amine activated beads
- Carboxy activated beads
- Epoxy activated beads
- Maleimide activated beads
- NHS activated beads
PureCube Epoxy Activated Beads in Research
Video of the Intelligent Infusion Masterclass: Thoman Bortecen about PureCube Epoxy Activated Beads using it for a semi-automated workflow for the quantitative analysis of the newly synthesized proteome.
Through the use of novel magnetic alkyne beads, the enrichment of newly synthesized proteins can be performed with a semi-automated protocol using 10-fold lower input.
Buy also at
Reach out to us
Anton Starke
Strategic Partnership Manager
Protein Purification
Anton.Starke@cube-biotech.com
+49-2173-99373-16
Sebastian Lühn
Strategic Partnership Manager
Protein Purification
Sebastian.Luehn@cube-biotech.com
+49-2173-99373-16