Figure 1. Flow chart of purification of DYKDDDDK-tagged protein

Figure 2. Flow chart of immunoprecipitation of DYKDDDDK-tagged protein

Lanes:
1. Eluate purified by GenScript resin
2. Eluate purified by Competitor-S resin
3. DYKDDDDK-tagged protein (~55 kDa)
4. Whole cell lysate
The red arrows indicate the bands of the target protein.

Figure 3. The DYKDDDDK-tagged proteins from cell lysates of various expression systems were captured by GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) and Competitor-S resin, respectively. The captured proteins were eluted by 0.4 mg/ml MDYKDHDGDYKDHDIDYKDDDDK peptide in TBS, and then were analyzed by SDS-PAGE. The results showed that GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) efficiently captured DYKDDDDK-tagged proteins from cell lysates of various expression systems. Especially, GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) significantly outperforms the competing product when purifying the target protein from yeast and E. coli lysates.

Figure 4. Purify the DYKDDDDK-tagged protein by GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907). The target protein was eluted by 0.1 mg/ml DYKDDDDK peptide in TBS. Then, the resin was regenerated by the regeneration solution for repeated use. The coefficient of variation (CV) of the amount of eluted target protein was less than 20% for 10 purification processes in total, indicating that GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) is capable of multiple repeated use with relatively high consistency when eluted by DYKDDDDK peptide.

Figure 5. Purify the DYKDDDDK-tagged protein by GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907). The protein was eluted by 0.4 mg/ml MDYKDHDGDYKDHDIDYKDDDDK peptide in TBS. Then, the resin was regenerated by the regeneration solution for repeated use. The coefficient of variation (CV) of the amount of eluted target protein was less than 20% among 9 purification processes in total, indicating that GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) is capable of multiple repeated use with relatively high consistency as well when eluted by MDYKDHDGDYKDHDIDYKDDDDK peptide.

Lanes:
1/3/5. IP Eluate by GenScript resin
2/4/6. IP Eluate by Competitor-S resin
7. DYKDDDDK-tagged protein (~55 kDa)
8. Whole HEK293 lysate

Figure 6. Different amounts (5/10/25 ng) of DYKDDDDK-tagged protein (~55 kDa) were spiked to 0.5 ml of HEK293 lysis buffer, and captured by GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) and Competitor-S resin, respectively. After washing step, the resin (containing the target protein) was then eluted by SDS-PAGE sample loading buffer, followed by a Western Blotting analysis of the eluted protein. The data showed that GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) outperforms the competing product in sensitivity for immunoprecipitation applications.

Lanes:
1. HA-tagged protein (~20 kDa)
2. Competitor-S resin
3. Competitor-T resin
4. Competitor-ST resin
5. GenScript resin

Figure 7. 10 μg of HA-tagged protein was incubated with GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No.L00907) and other competitor products, respectively. The proteins were eluted by SDS-PAGE sample loading buffer, then analyzed by Western blotting. Data showed that GenScript Anti-DYKDDDDK Affinity Resin Easy (Cat. No. L00907) exhibited extremely low nonspecific binding to the HA-tagged protein and outperforms the competing products.