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Anti-DYKDDDDK G1 Affinity Resin

Anti-DYKDDDDK G1 Affinity Resin

Figure 1.
Purification of DYKDDDDK‐tagged protein from E. coli lysate

Lanes:
1. Whole cell lysate
2. Flow through
3. Protein marker
4‐8. Eluate of target DYKDDDDK‐tagged protein (M.W. 60.4 kDa)

Anti-DYKDDDDK G1 Affinity Resin

Figure 2.
Purification of N‐ terminus and Cterminus DYKDDDDK ‐tagged proteins

Lanes:
1‐2．Eluate of N‐terminus DYKDDDDKtagged protein (M.W. 19.2 kDa)
3. Protein marker
4‐5. Eluate of C‐terminus DYKDDDDKtagged protein (M.W. 26.4 kDa)

Anti-DYKDDDDK G1 Affinity Resin

Figure 3.
Purification of DYKDDDDK‐tagged protein from mammalian cells

Lanes:
1. Protein marker
2. Supernatant from cultured CHO cells
3. Flow through after binding
4. Eluate of target DYKDDDDK ‐tagged protein (M.W. 27.6 kDa)

Anti-DYKDDDDK G1 Affinity Resin

Figure 4.
Purification of DYKDDDDK‐tagged glycoprotein from mammalian cells

The upper figure shows SDS‐PAGE analysis result and the lower figure shows Western blot result.

Lanes:
1. Protein marker
2. Supernatant from CHO cells expressing target glycoprotein
3. Flow through after binding
4. Eluate of target DYKDDDDK‐tagged protein (M.W. 50.0 kDa)

Anti-DYKDDDDK G1 Affinity Resin

Figure 5.
Purification of DYKDDDDK‐tagged proteins using different elution methods.

Lanes:
1. Protein #1 (M.W. 16.8 kDa) by competitive elution using 300 μg/ml DYKDDDDK peptide.
2. Protein #2 (M.W. 54.6 kDa) by neutral elution with 3M NaCl
3. Protein #3 (M.W. 27.6 kDa) by acid elution, pH 3.5
4. Protein #1 (M.W. 16.8 kDa) by alkaline elution, pH 10.5
5. Protein #1 (M.W. 16.8 kDa) by alkaline elution, pH 12

Anti-DYKDDDDK G1 Affinity Resin

Figure 6.
Benchmark with Sigma‐Aldrich’s ANTI‐FLAG M2 Affinity Gel

Side‐by‐side comparison of DYKDDDDKtagged protein purification with GenScript’s Anti‐DYKDDDDK G1 Affinity Resin and Sigma‐Aldrich’s ANTI‐FLAG M2 Affinity Gel, from same staring material following manufacturer suggested procedures.

Lanes:
1. Protein marker
2. Purification from M2 gel (pH 3.5 elution)
3. Purification from G1 resin (pH 12.0 elution)

Anti-DYKDDDDK G1 Affinity Resin

GenScript Anti‐DYKDDDDK G1 Affinity Resin (Cat. No. L00432) is designed for the purification of DYKDDDDK‐tagged protein from commonly used protein expression systems including bacteria, yeast and mammalian cells.
L00432
¥40,600.00

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Overview
Product Description DYKDDDDK or FLAG® 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. GenScript Anti‐DYKDDDDK G1 Affinity Resin (Cat. No. L00432) is designed for the purification of DYKDDDDK‐tagged protein from commonly used protein expression systems including bacteria, yeast and mammalian cells. The DYKDDDDK‐tagged protein in the cell lysate can specifically bind to the anti‐DYKDDDDK monoclonal antibodies coupled to the resin. Nonspecific bindings are eliminated by stringent washing steps, and the target protein is finally eluted with high recovery rate. 
Key Features Comparison between Anti-DYKDDDDK IP Resin and Anti-DYKDDDDK G1 Affinity Resin.
Anti-DYKDDDDK G1 Affinity Resin (L00432) Anti-DYKDDDDK IP Resin (L00425)
Immunoprecipitation (IP) √ √
Purification √ n/a
Elution Alkaline elution √ n/a
Alkalescent elution √ n/a
Acid elution √ n/a
Neutral elution √ n/a
Competitive elution √ n/a
SDS-PAGE loading synthesize buffer √ √
Regeneration 4-10 times not recommend
Binding volume large scale mini-volume (<1ml)

Properties
Product Content 50% settled resin in TBS with 0.02% sodium azide
Matrix 4% cross‐linked agarose
Average Bead size μm
Ligand Mouse monoclonal antibody against DYKDDDDK tag, clone G1  
Binding Capacity Approximately 1.5 mg DYKDDDDK‐tagged protein per ml settled resin
Storage & Stability Store at 2‐8 °C for up to 12 months. Do not freeze the resin.
Resin Reuse The resin can be recycled for at least 4 times. If maintained properly, the resin can be reused 10 times with minimum loss of binding capability.
Elution Method Acid, alkaline, neutral, peptide competitive or SDS‐PAGE loading buffer elutions*
(*If elute with SDS-PAGE loading buffer, the heavy chain of G1 antibody will be denatured and release from the resin.)
Reagents Compatibility Compatible with commonly used reagents at certain concentrations

Examples
  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 1.
    Purification of DYKDDDDK‐tagged protein from E. coli lysate

    Lanes:
    1. Whole cell lysate
    2. Flow through
    3. Protein marker
    4‐8. Eluate of target DYKDDDDK‐tagged protein (M.W. 60.4 kDa)

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 2.
    Purification of N‐ terminus and Cterminus DYKDDDDK ‐tagged proteins

    Lanes:
    1‐2．Eluate of N‐terminus DYKDDDDKtagged protein (M.W. 19.2 kDa)
    3. Protein marker
    4‐5. Eluate of C‐terminus DYKDDDDKtagged protein (M.W. 26.4 kDa)

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 3.
    Purification of DYKDDDDK‐tagged protein from mammalian cells

    Lanes:
    1. Protein marker
    2. Supernatant from cultured CHO cells
    3. Flow through after binding
    4. Eluate of target DYKDDDDK ‐tagged protein (M.W. 27.6 kDa)

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 4.
    Purification of DYKDDDDK‐tagged glycoprotein from mammalian cells

    The upper figure shows SDS‐PAGE analysis result and the lower figure shows Western blot result.

    Lanes:
    1. Protein marker
    2. Supernatant from CHO cells expressing target glycoprotein
    3. Flow through after binding
    4. Eluate of target DYKDDDDK‐tagged protein (M.W. 50.0 kDa)

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 5.
    Purification of DYKDDDDK‐tagged proteins using different elution methods.

    Lanes:
    1. Protein #1 (M.W. 16.8 kDa) by competitive elution using 300 μg/ml DYKDDDDK peptide.
    2. Protein #2 (M.W. 54.6 kDa) by neutral elution with 3M NaCl
    3. Protein #3 (M.W. 27.6 kDa) by acid elution, pH 3.5
    4. Protein #1 (M.W. 16.8 kDa) by alkaline elution, pH 10.5
    5. Protein #1 (M.W. 16.8 kDa) by alkaline elution, pH 12

  • Anti-DYKDDDDK G1 Affinity Resin
    • Anti-DYKDDDDK G1 Affinity Resin

    Figure 6.
    Benchmark with Sigma‐Aldrich’s ANTI‐FLAG M2 Affinity Gel

    Side‐by‐side comparison of DYKDDDDKtagged protein purification with GenScript’s Anti‐DYKDDDDK G1 Affinity Resin and Sigma‐Aldrich’s ANTI‐FLAG M2 Affinity Gel, from same staring material following manufacturer suggested procedures.

    Lanes:
    1. Protein marker
    2. Purification from M2 gel (pH 3.5 elution)
    3. Purification from G1 resin (pH 12.0 elution)