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MonoRab™ Anti-DYKDDDDK Affinity Resin

GenScript’s MonoRabᵀᴹ Anti-DYKDDDDK Affinity Resin is designed for optimal purification of DYKDDDDK-tagged proteins. DYKDDDDK tag is an octapeptide tag that can be added to a protein using recombinant DNA technology. It can be fused to the N-terminus or C-terminus of a target protein to facilitate detection and purification. Based on our new rabbit monoclonal antibody technology (MonoRabᵀᴹ), this resin offers high binding capacity, high purity, and resistance to high concentrations of salt or acid during protein purification. The binding capacity of MonoRabᵀᴹ 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.
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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. Table 1 lists the main characteristics of the MonoRab™ Anti-DYKDDDDK Affinity Resin.
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Product Content 50% settled resin in TBS with 50% glycerol and 0.02% sodium azide
Matrix 4% cross-linked agarose
Average Bead size 90 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*.
Resin Reuse 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
Storage & Stability Store at -20 °C for 24 months.
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MonoRab™ Anti-DYKDDDDK Affinity Resin

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MonoRab™ Anti-DYKDDDDK Affinity Resin

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) »

MonoRab™ Anti-DYKDDDDK Affinity Resin

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) »

MonoRab™ Anti-DYKDDDDK Affinity Resin

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) »

MonoRab™ Anti-DYKDDDDK Affinity Resin

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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 For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.