3X FLAG Peptide for Elution

Overview

Tested applications

For use in competitive elution of 3X FLAG® fusion proteins from the anti-FLAG monoclonal antibody (Cat no: AT0022) in solution or bound to agarose beads on the anti-FLAG agarose affinity gel (Cat no: AT0078). 

Product Picture

Properties

Sequence	Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys
Sequence name	3X FLAG Peptide
Molecular Formula	C120H169N31O49S
Molecular Weight	2861.87
Purity	> 95%
Storage	Store at -20°C.

Working Concentration	0.1mg-1 mg/mL Note: In order to obtain best results in different techniques we recommend determining optimal working dilution by titration test.
Form	lyophilized powder
Preparation of stock solution	To prepare a stock solution, dissolve in TBS (50 mMTris-HCl, pH 7.4, with 150 mM NaCl) at a concentration of 5 mg/ml.

 

Applications

Protocols

Elution protocol with 3X FLAG peptide
Prepare 3X FLAG peptide stock solution (5 mg/mL). 

Dissolve 3X FLAG peptide in TBS (50mM Tris HCL, 150 mM NaCl, pH7.4) to a final concentration of 5 mg/mL. For extended storage after reconstitution, store at –20 °C in with 50% glycerol. Avoid repeated freeze-thaw.

 

2. Prepare 3X FLAG peptide working solution (1 mg/mL).

Dilute 5-fold with TBS to prepare a 3X FLAG peptide working solution containing 1 mg/mL of 3X FLAG peptide. 

 

3. Add 50 µL of 3X FLAG peptide working solution into each immuno complex of the protein and anti FLAG beads (initially adding 40 μL beads slurry)

 

4. Gently mix and incubate the samples and controls at 37 ℃ on a rotator for 20 minutes.

 

5. Centrifuge the agarose beads for 60 seconds at 8,000×g. Transfer the supernatants into fresh EP tubes. 

Or

Separate the magnetic beads on a magnetic separation rack and save the supernatant containing the target antigen protein.

Be careful not to transfer any beads.

 

6. Repeat elution step once for higher yields.