Poly DYKDDDDK Tag Peptide lyophilized powder

Selleck's Poly FLAG Peptide lyophilized powder Has Been Cited by 19 Publications

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Description

Poly DYKDDDDK Tag Peptide lyophilized powder(3X DYKDDDDK Tag Peptide) is a polypeptide composed of 23 amino acids with a molecular weight of 2864 Da, which can bind Anti-DYKDDDDK Tag antibody through competition, so that the antibody bound to Anti-DYKDDDDK Tag antibody can be eluted during immunoprecipitation DYKDDDDK Tag fusion expression protein. The motif Asp-Tyr-Lys-Xaa-Xaa-Asp in the polypeptide is repeated three times, and the 8 amino acids at the carbon-terminus constitute the classic DYKDDDDK Tag (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys).

Properties

Appearance (Color)	White
Appearance (Form)	Powder
Form	Lyophilized Powder
Purity by HPLC-MS	95.33 % (the latest lot) Poly DYKDDDDK Tag Peptide lyophilized powder
Concentration	Recommended working concentration is 200-400 μg/mL for elute DYKDDDDK Tag fusion proteins from the Anti-DYKDDDDK beads.
Storage Temperature	2-8°C

Application

For use in competitive elution of DYKDDDDK fusion proteins from the ANTI-DYKDDDDK Tag monoclonal antibody in solution or bound to agarose on the Anti-DYKDDDDK 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 Fusion Protein from Anti-DYKDDDDK beads: 1. Thoroughly suspend the Anti-DYKDDDDK Tag 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-DYKDDDDK Tag 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 Fusion Protein by Competition with Poly DYKDDDDK Peptide. Elute the bound DYKDDDDK Fusion Proteinby competitive elution with five one-column volume aliquots of a solution containing 200-400 ug/mL Poly DYKDDDDK Peptide in TBS.; 8. Recycling the Anti-DYKDDDDK beads
Poly DYKDDDDK Peptidemay not elute all of the DYKDDDDK Fusion Protein bound to Anti-DYKDDDDK beads. It is recommended the Anti-DYKDDDDK 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 beads in glycine HCl for longer than 20 minutes.
9. Storing the Anti-DYKDDDDK beads Wash the Anti-DYKDDDDK 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.

Poly DYKDDDDK Tag Peptide lyophilized powder

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Description

Properties

Application

Attention

Procedure

Storage (From the date of receipt)

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