Bimake/Poly FLAG Peptide lyophilized powder/4 mg/B23111

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Description

The Poly DYKDDDDK (FLAG) Peptide lyophilized powder was synthesized by 23 amino acid residues with molecular weight 2,864 Da. The Asp-Tyr-Lys-Xaa-Xaa-Asp motif is repeated three times in the peptide. Eight amino acids at the C-terminus make up the classic FLAG sequence (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys).

Properties

Application

For use in competitive elution of DYKDDDDK (FLAG) fusion proteins from the ANTI-FLAGmonoclonal antibody in solution or bound to agarose on the Anti-DYKDDDDK (FLAG) beads.

Attention

1. Read the User Manual carefully before the first use; 2. Avoid freezing, drying and high-speed centrifugation during use and storage of beads; 3. This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

Procedure

Peptide Elution of DYKDDDDK(FLAG) Fusion Protein from Anti-DYKDDDDK(FLAG) beads

1. Thoroughly suspend the Anti-Flag Affinity Gel in the vial, for a uniform suspension of the resin. Quickly transfer 10μl of the gel suspension (about 5μl of packed gel volume) to a fresh tube.

2. Add 0.6 mL TBS. Thoroughly suspend the Anti-Flag Affinity Gel by pipetting. Centrifuge the resin at 5000 rpm for 30 secondsand carefully remove the supernatant. Be sure to remove all of the wash buffer without discarding the resin. Repeat 3-4 times.

3. Add 500 μL of cell periplasmic extracts to the washed resin.

4. Gently agitate samples for 2 hours at 4°C. 5. Centrifuge the resin for 30 seconds at 5000 rpm. Transfer the supernatants to a fresh tube. 6. Wash the resin with 0.5mL TBS until the OD280 of the supernatant reads＜0.05. 7. Elution of DYKDDDDK (FLAG) Fusion Protein by Competition with Poly DYKDDDDK (FLAG) Peptide. Elute the bound DYKDDDDK(FLAG) Fusion Proteinby competitive elution with five one-column volume aliquots of a solution containing 200-400 ug/mL Poly DYKDDDDK(FLAG) Peptide in TBS.8. Recycling the Anti-DYKDDDDK(FLAG) beads Poly DYKDDDDK(FLAG) Peptidemay not elute all of the DYKDDDDK(FLAG) Fusion Protein bound to Anti-DYKDDDDK(FLAG) beads. It is recommended the Anti-DYKDDDDK(FLAG) beads be regenerated immediately after use by washing with three 5 mL aliquots of 0.1 M glycine HCl, pH 3.5. The gel should be immediately re-equilibrated in TBS until the effluent is at neutral pH. Note: Do not leave the Anti-DYKDDDDK(FLAG) beads in glycine HCl for longer than 20 minutes. 9. Storing the Anti-DYKDDDDK(FLAG) beads Wash the Anti-DYKDDDDK(FLAG) beads three times with 5 mL of 50% glycerol with 10mM sodium phosphate, 150 mM sodium chloride, pH 7.4, containing 0.02% (w/v) sodium azide. then add another 5 mL of 50% glycerol with 10 mM sodium phosphate, 150 mM sodium chloride, pH 7.4, containing 0.02% (w/v) sodium azideand store at -20°C without draining.

Storage & stability

Store the product at 2-8°C.

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Poly FLAG Peptide lyophilized powder