G BOND Ni-IDA Agarose Bead
Protein Extraction Buffer
Protein Extraction Buffer delivers high amount of Protein from minimum amount of sample source
- GSure™ DNA Extraction & Purification Kit
- GSure™ DNA Extraction & Purification Reagents
- Gel Loading Dyes
- DNA Ladder
- Gel Stain
- Running Buffer
- Agarose
Find support for technicalities of applications and product use
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Applications
This GBond Ni-IDA based purified protein can be reliably used in applications like
- For structural or functional assay
- For antibody production
- DNA- Protein interaction
The cost-effective IDA products for His-tag protein purification by gravity-flow
| Technology | IMAC (immobilized metal ion affinity chromatography) |
| Chelating ligand | IDA (iminodiacetic acid) |
| Supplied | 50 % (v/v) aqueous suspension containing 20 % ethanol |
| Matrix | 6 % beaded agarose (cross-linked |
| Binding capacity* | precharged with Ni2+up to 50 mg/mL |
| Storage temperature | settled agarose 4–8 °C |
Decision to purify 6xHis-tagged proteins:
- Native conditions: Protein is soluble (in the supernatant after lysis) and you want to preserve protein activity.
- Denaturing conditions: protein is insoluble (in the pellet after lysis) or protein activity is not important for downstream
Ordering Information
| Cat. No. | Product | Pack Size | Price |
| GBI-01A | G Bond Ni-IDA agarose bead | 10 prep | POR |
| GBI-01B | G Bond Ni-IDA agarose bead | 25 prep | POR |
| GBI-01C | G Bond Ni-IDA agarose bead | 50 prep | POR |
| GBI-01D | G Bond Ni-IDA agarose bead | 100 prep | POR |
Product Details Specifications Protocols & Manual Product Resource
Description
MAC resin was first developed by Porath and colleagues in 1975 by fixing metal ions to agarose with iminodiacetic acid (IDA). Since then this ligand is widely used as commonly used commercial IMAC resins along with NTA resin for affinity purification of proteins. Structurally, IDA and NTA differ by the presence of an additional carboxymethyl group on NTA. Chemically, the additional functional group makes NTA a stronger coordinator of metal ions because it has 4 valencies available compared to the 3 of IDA. While in NTA ligands have 2 valences available to interact with the imidazole rings of histidine residues, in IDA ligands have 3 valences for such interaction. Thus the coordination number plays a vital role in the quality of the resulting purified protein fractions. The tridentate ligand requires a lower imidazole concentration to elute protein then tetradentate NTA. Lastly, IDA is a smaller molecule which can be coupled to the matrix at a higher density resulting in a higher metal loading capacity. Generally, low loading density enhances the quality of the purification but results in lower target protein recovery, whereas high loading density generates greater yields but with higher unspecific binding. In such case an optimization of protein: Ligand ratio can give the best results in quality as well as in quantity. Lastly, resins constructed with this ligand are generally less expensive.
G BOND Ni-IDA Agarose Bead
Product Details
Ni-IDA matrix is an affinity chromatography for purifying recombinant 6xHis-tagged proteins expressed in bacteria, insect, and mammalian cells. This matrix offers high binding capacity and minimal nonspecific binding under optimized condition. It can be used in both batch and column based purification chromatography. The resin comes in 50% slurry in 20% ethanol. Cleared cell lysates are loaded onto the matrix. His-tagged proteins are bound, and other proteins pass through the matrix. After washing, Histagged proteins bound to the resin are eluted with low pH buffer or by competition with imidazole or histidine under native or denaturing conditions. The resulting proteins are ready for use in target applications.
NOTE: Proteins may be purified on Ni-IDA resins in either a batch or a column procedure. The batch procedure entails binding the protein to the Ni-IDA resin in solution and then packing the protein–resin complex into a column for the washing and elution steps. This strategy promotes efficient binding of the 6xHistagged protein especially when the 6xHis tag is not fully accessible or when the protein in the lysate is present at a very low concentration. In the column procedure, the Ni-IDA resin is first packed into the column and equilibrated with the lysis buffer. The cell lysate is then slowly applied to the column. Washing and elution steps are identical in the batch and column procedure.
Protocol & Manuals
Some product description and more information to download our product brochure or manual datasheet
| Catalogue No. | Product Details | Datasheet | Brochure |
| GBI-01A | G Bond Ni-IDA agarose bead, 10 ml |  |
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| GBI-01B | G Bond Ni-IDA agarose bead, 25 ml | |
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| GBI-01C | G Bond Ni-IDA agarose bead, 50 ml | |
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| GBI-01D | G Bond Ni-IDA agarose bead, 100ml | |
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