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| L00766-1 | |
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| Product description | DYKDDDDK or FLAG® tag is an octapeptide that can be added to a protein using recombinant DNA technology. The tag sequence can be fused to the coding sequence of a target protein to facilitate detection and purification. GenScript’s MonoRab™ Anti-DYKDDDDK Affinity Resin (Cat. No. L00766) is designed for optimal purification of DYKDDDDK-tagged proteins. Based on our new rabbit monoclonal antibody technology (MonoRab™), this resin offers high binding capacity, high purity, and resistance to high concentrations of salt or acid. The binding capacity of MonoRab™ Anti-DYKDDDDK Affinity Resin is 1.5 mg DYKDDDDK-tagged protein per mL of settled resin. The recombinant proteins expressed in bacterial or mammalian cells cultures specifically bind to the anti-DYKDDDDK monoclonal antibodies coupled to the resin, resulting in highly pure eluates. The resin can withstand stringent washing steps, especially with buffers containing high concentration of salt. The resin is resistant to up to 2 M NaCl, and to buffers with pH as low as 2.5. |
| Key Features |
● High binding capacity: 1.5 mg DYKDDDDK-tagged protein per mL of settled resin ● High purity ● Resistance to high concentrations of salt (up to 2 M NaCl) ● Resistance to low pH buffers (as low as 2.5) |
| Formulation | 50% settled resin in TBS with 50% glycerol and 0.02% sodium azide |
| Matrix | 4% cross-linked agarose |
| Average Particle Size | 90 μm (45-165 μm) |
| Ligand | MonoRab™ monoclonal antibody against DYKDDDDK tag |
| Binding Capacity | Approximately 1.5 mg DYKDDDDK-tagged protein (Size: 50KDa) per ml settled resin |
| Elution Method |
Two methods can be implied for elution: 1. Alkaline, Elution under native conditions by competition with peptide 2. Elution with SDS-PAGE loading buffer* *When eluting with SDS-PAGE loading buffer, the heavy chain of the MonoRabTM antibody will be denatured and released from the resin. |
| Reuse Capability | The resin can be recycled for at least 4 times. If maintained as per manufacturer’s instructions, the resin can be reused 10 times with minimum loss of binding capability. |
| Reagents Compatibility | Compatible with commonly used reagents at certain concentrations |
| Application | purification of DYKDDDDK-tagged proteins |
| Storage | Store at -20 °C. |
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Figure 1. Purification of DYKDDDDK-tagged protein from E. coli lysate and elution with alkaline elution buffer , pH 12.0
Lanes:
M. Protein Marker
1. Whole cell lysate
2. Flow through
3-7. Elution fractions of target DYKDDDDK-tagged protein (M.W. 50.4 kDa)
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Figure 2. Purification of DYKDDDDK-tagged protein from E. coli lysate and Elution with 300 μg/ml DYKDDDDK peptide
Lanes:
M. Protein Marker
1. Whole cell lysate
2. Flow through
3-7. Elution fractions of target DYKDDDDK-tagged protein (M.W. 50.4 kDa)
»
Figure 3.
Purification of DYKDDDDK-tagged protein from mammalian cell lysate and elution with alkaline elution buffer , pH 12.0
Lanes:
M. Protein Marker
1. Whole cell lysate
2. Flow through
3-7. Elution fractions of target DYKDDDDK-tagged protein (M.W. 50.4 kDa)
»
Figure 4.
Benchmark with competitor’s ANTI-FLAG Affinity Gel
Side-by-side comparison of DYKDDDDK-tagged protein purification with GenScript’s MonoRabTM Anti-DYKDDDDK Affinity Resin and competitor’s ANTI-FLAG® Affinity Gel, from same staring material and incubated in 2 M NaCl containing buffer, following with manufacturer suggested procedures.
Lanes:
M. Protein Marker
1. Whole cell lysate
2-4. Purified with GenScript’s MonoRabTM Anti-DYKDDDDK Affinity Resin
5-7. Purified with competitor’s ANTI-FLAG® Affinity Gel
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| Terms of Use | This product may be the subject of one or more patents filed by GenScript Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer may not sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for any commercial purposes. For commercial use, please contact GenScript at product@genscript.com. |
For research and manufacturing use. Direct human use, including taking orally and injection are forbidden.
