Anti-DYKDDDDK Affinity Resin

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Introduction

Anti-DYKDDDDK-tag agarose is a mouse IgG1 monoclonal antibody covalently coupled on agarose. It is used for purification or immunoprecipitation of DYKDDDDK-tag fusion proteins.

Parameter Information
Antibody High-affinity mouse IgG1 monoclonal Anti- DYKDDDDK-tag antibody
Agarose 4% cross-linked agarose beads
Particle diameter The particle size range is 45-165 μm, and the average particle size is 90 μm
Capacity ≥0.5mg/mL flag resin (70kDa fusion protein)
Binding Specificity The antibody can bind to DYKDDDDK-tag at N-terminal, Met-N-terminal, C-terminal, and internal locations of a DYKDDDDK-tag fusion protein
Product status Suspension medium, bead is white, solution is colorless
Velocity 0.5-1.0 mL/min is recommended for 1mL column
Pressure ≤ 0.3 MPa

 

Product Features

1. More stable and higher binding capacity than competitor's;
2. Suitable for both N-terminal and C-terminal tagged proteins;
3. Suitable for viscous samples
4. Can be used in a column chromatography, batch Chromatography and immunoprecipitation
5. Spot stocks with strict quality control

Shipping:

Shipped at ambient temperature. Please store it at 2~8℃or –20℃

Storage:

The unopened product is stable for one year from the date of receipt when stored at 2~8℃. After use, the resin should be cleaned and stored in PBS buffer containing 0.03% (v/v) ProClin 300 at 2~8℃. Or add 50% glycerol with PBS buffer containing 0.03% (v/v) ProClin 300 store at -20℃. Avoid repeated freezing and thawing. Do not freeze without glycerol.

Formulation:

Anti-DYKDDDDK-tag agarose is supplied as a 50% slurry in PBS (10 mM Na2HPO4, 2.7 mM K2HPO4, 137 mM NaCl, pH 7.4) containing 0.03% (v/v) ProClin 300.

Reagents Required but not Provided:

Binding Buffer: PBS (10 mM phosphate, 2.7 mM potassium chloride, and 137 mM sodium chloride, pH 7.4)
Elution Buffer: 100 mM Glycine, 10 mM NaCl, pH 3.0
Neutralization Buffer: 1M Tris-HCl, pH8-9

Experimental Data

Fig.1. Binding capacity comparison between a competitor's (A) and Sinobiological's resin(B):The competitor's (A) and Sinobiological's resin(B) were incubated with the same amount of DYKDDDDK tagged protein(>2 mg, excessive)and purified according to the instruction respectively. The binding capacity was shown using SDS-PAGE (13%) and UV-Vis.

Fig.2.Suitable for both N-terminal and C-terminal tagged proteins:Sinobiological's resin was used in the purification of sample with protein A\B\C\D\E (C-terminal tagged) and protein F (N-terminal tagged). The purification result was shown using SDS-PAGE (13%) and UV-Vis.

Fig.3.Can be regenerated many cycles for low viscous sample purification:The competitor's (A) and Sinobiological's resin(B) (1mL) were used in the purification of low viscous sample(20mL).Both of the resin can be regenerated up to 20 cycles without obvious binding capacity change.The binding capacity was shown using SDS-PAGE (13%) and UV-Vis.

Fig.4.Suitable for repeated usage of high viscous samples' purification:1mL of the Sinobiological's resin was used in the purification of the high viscous sample(20mL) to get the target protein(about 40kD, C-terminal DYKDDDDK tagged) for 6 times. The binding capacity was checked by 13% SDS-PAGE and UV-Vis.

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