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Customized resins - maximum specificity


What are custom beads for?

If commonly used protein tags do not work for your protein purification, you do not have to despair. The solution to your problem is the use of protein specific ligands instead of artificially added tags. These ligands are the key for 100% specific purification of the desired protein. This way purification from native tissue as well as from denaturing enviroments are possible.

Cube Biotech has performed many custom resin & beads projects and you can make this experience work for you!
One of our numerous successful examples (Table 1) is the coupling of the cobra venom neurotoxin-3 onto agarose resin. This leads to a high-yield and 100% specific purification of the Acetylcholine receptor (AChR) Torpedo california, which the venom binds to naturally. In fact this project was so successfull that we added AChR to our standard product catalogue!

 



Examples for Custom Beads by Cube Biotech

overview custom resins
Fig. 1: Overview of some of the molecules and their respective classes we have already successful coupled to either our agarose resins or MagBeads. All combination led to high quality protein purifications.
The applications of customized resins/beads extent to numerous possibility of proteomics and sometimes beyond. Figure 1 shows our favourite example of custom beads, while more is listed in table 1.
Table 1: Customized Resins from Cube Biotech. Click on the arrow to see the full list.
 


Custom bead creation

Three decisions have to be made for the creation of a customized bead product by Cube Biotech. Table 2 leads you through these decision process and gives you some guidance.
Table 2: Choose matrix, format and ligand and we will create your custom resin.
1. Choose a matrix
The first question that need to be answered is "Agarose resin or MagBeads?" This is mostly a question on your personal preference to on of the methods. Do you favor batch spin columns and drip columns or do you prefer magnetic bead based methods?
Agarose Resin Magnetic Beads
agarose scheme
Our agarose resin is very homogeneous in size, yielding a high degree of reproducibility between individual purification runs. The cross-linked agarose is physically stable and highly porous for optimal protein interaction.
magnetic agarose
Our ferrimagnetic agarose beads are produced in house and are ideal for extracting proteins from dilute samples or for pull-down experiments.
2. Choose a format
The second decision is regarding the format in which the resin or MagBeads should be created. For MagBeads bulk deliveries are preferred only. While in regards to agarose resin you can choose between bulk resin and pre-packed resin in FPLC Columns.
Bulk FPLC Columns
  • Bulk resins are delivered as a 50% suspension in a buffer containing 20% ethanol. Various volumes are available.
  • MagBeads are usually delivered as a 25% suspension in a buffer containing 20% ethanol. Various volumes are available.
Resins are prepacked in high-quality columns of 1 or 5 mL bed volume, ready to be used on most FPLC workstations.
3. Choose a tag or ligand
The third and probably the most important question surrounds the ligand that should be coupled to the beads. Have a look at table 1 to see what we already managed to accomplish.  
Nonetheless you should have a look our standard protein purification products, maybe one of you existing beads can help you out.
Cube Biotech protein purification products
Polyhistidine / His Tag
The workhorse of affinity tags. This His Tag is short and can be added to both termini of a protein. The tag is designed for highest protein yields.
Our His affinity agarose resins and MagBeads by Cube Biotech for highest protein yields.
Glutathion Affinity / GST Tag
A 26 kDA large affinity tag that consists of the protein Glutathione-S-transferase. Preferably added to the N-Terminus
Our GST Tag affinity agarose resins and MagBeads by Cube Biotech for a great balance between protein yield and purification specificity.
Streptactin / Strep-tag®
An eight amino acid long affinity tag that provides high levels of protein purity
Our Strep-tag® affinity products by Cube Biotech as a smaller alternative to the GST tag while having similar purification properties.
Rho1D4 Tag
A nine amino acid long affinity tag that provides our highest levels of protein purity and membrane protein purification
Our Rho1D4 Tag affinity products by Cube Biotech for membrane protein purification.
Hydrophobic Interaction resins
A reasonable option for protein purification using hydrophobic interactions between protein and agarose bead.
Our HIC products by Cube Biotech protein purification without the use of affinity tags.
Your ligand
Protein-specific ligands bind targets in particular oligomeric states, conformations or orientations, enabling specialized applications.
PureCube Tailor-made Resins
Our custom resins are produced within a couple weeks after ordering and/or consultation with our chemistry experts. You lay out the matrix parameters (e.g., ligands, linker, spacer, etc.), and we produce it in the quantities you need.
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