In the field of biomedical research, protein purification is a key technology for exploring the mysteries of life. However, traditional purification methods struggle with inefficiency and high costs—especially for membrane proteins, which are the focus of structural biology research. These proteins are structurally complex, have low expression levels in cell membranes, and are sensitive to environmental conditions, making their purification particularly difficult. Now, Yeasen Biotech, with its outstanding R&D capabilities, has launched the upgraded Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES), a powerful tool for efficient and convenient protein purification, even for challenging membrane proteins.
Product Performance
1. High Specificity and High Binding Capacity: Key Advantages for Target Protein Capture
Yeasen Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES) is prepared by covalently coupling high-quality mouse IgG2b monoclonal antibodies with 4% highly cross-linked agarose gel. This unique manufacturing process endows the product with extremely high specificity, enabling it to precisely recognize and bind fusion proteins with the Flag tag. Whether it is the N-terminal Flag fusion protein modified by Met (Met-Flag-Protein), the N-terminal Flag fusion protein (Flag-Protein), or the C-terminal Flag fusion protein (Protein-Flag), the gel can handle them with ease.
In membrane protein purification, this specificity is particularly important because these proteins are structurally complex and prone to non-specific binding with other proteins. The high specificity of the Flag gel effectively reduces interference from non-specific proteins, ensuring high purity of the purified membrane proteins. Additionally, the gel has a high binding capacity for Flag-tagged fusion proteins (at least 1.1 mg protein/mL gel). This means that even if the target protein is expressed at low levels, it can still be efficiently captured and purified using the Flag gel.
2. High Purity and High Yield: Enabling Downstream Applications
Yeasen Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES) has minimal non-specific binding of impurities, resulting in highly pure protein samples after purification. This is crucial for downstream applications such as bioactivity assays and structural analysis. High-purity protein samples provide more accurate experimental data, laying a solid foundation for researchers' studies.
3. Optimized Experimental Workflow: Simplifying Protein Purification Challenges
Protein purification needs to be conducted under mild conditions to avoid structural damage to the proteins, especially for complex membrane proteins. Yeasen Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES) is designed with this in mind. Its storage buffer is TBS containing 50% glycerol at pH 7.4, which not only protects the activity of the gel but also provides a gentle environment for protein purification.
The purification process with the Flag gel is simple and efficient. To facilitate the use of 1×Flag peptide elution, the product package includes a complimentary 1×Flag peptide (Cat#20572ES). This makes the elution process gentle (with almost no damage to the proteins) and highly convenient (further enhancing the ease of experimentation).
4. Multiple Sizes and Flexible Shipping Options: Meeting Diverse Experimental Needs
Considering the varying scales and needs of different laboratories, Yeasen offers the Flag gel in multiple sizes, including 1 mL, 5 mL, 25 mL, and 100 mL. Whether for small-scale preliminary exploration or large-scale protein production, there is a suitable option available. For large-volume annual purchases, customers can choose to receive shipments in batches, allowing them to enjoy price discounts while avoiding the pressure of excessive inventory.
Test Data
 |
M: Protein Marker 1: Pre-column crude sample of Flag-N/ Flag-M/Flag-C protein 2: Post-column flow-through of Flag-N/ Flag-M/Flag-C protein 3: Elution (pH 2.7 elution) 4: Elution (pH 12 elution) 5: Elution (1×Flag peptide elution) 6: Purified gel (pH 2.7 elution) 7: Purified gel (pH 12 elution) 8: Purified gel (1×Flag peptide elution) a：Flag-N b：Flag-M c：Flag-C |
Figure 1. Yeasen Anti-DYKDDDDK(Flag) Affinity Gel purification results for low-expressing proteins (Flag-N (a), Flag-M (b), Flag-C (c)) shown in SDS-PAGE.
 |
M: Protein Marker 1: Pre-column crude sample of Flag-C protein 2: Post-column flow-through of Flag-C protein 3: Elution (pH 2.7 elution) 4: Elution (pH 12 elution) 5: Elution (1×Flag peptide elution) 6: Purified gel (pH 2.7 elution) 7: Purified gel (pH 12 elution) 8: Purified gel (1×Flag peptide elution) a：Yeasen b：Brand S* c：Brand G* |
Figure 2. Comparison of Yeasen (a) withBrand S* (b) and Brand G* (c) Anti-Flag gels in purifying low-expressing proteins (e.g., membrane proteins) shown in SDS-PAGE.
Conclusions:
1. Yeasen Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES) has high specificity and can bind to Flag tags in various positions (N-terminal Flag fusion proteins, Met-modified N-terminal Flag fusion proteins, C-terminal Flag fusion proteins).
2. Under conditions of extremely low target protein expression (e.g., membrane proteins), Yeasen Anti-DYKDDDDK(Flag) Affinity Gel (Cat#20584ES) achieves significantly higher yields and purities compared to competitors. The use of Yeasen complimentary 1×Flag peptide (Cat#20572ES) for elution provides even better results.
Complete Solutions for Flag-Tagged Fusion Proteins:
Recombinant Enterokinase (Cat#20395ES): A specific protease that cleaves the carboxyl end of lysine preceded by four aspartic acids: Asp-Asp-Asp-Asp-Lys. This sequence is part of the octapeptide Flag tag (DYKDDDDK). Recombinant Enterokinase (rEK) can easily remove the Flag tag from fusion proteins, making it an ideal tool for studying the structure and function of native proteins.
Anti-DYKDDDDK(Flag) MagBeads (Cat#20565ES): Polymer magnetic microspheres prepared by covalently coupling high-quality mouse anti-DYKDDDDK (Flag) monoclonal antibodies with silica-based magnetic beads (diameter of 200 nm). These beads are suitable for immunoprecipitation (IP) or co-immunoprecipitation (Co-IP) of proteins with the Flag tag.
Product Information
|
Product Name |
Product Number |
Specification |
|
1 mL/5 mL/25 mL |
||
|
Recombinant Bovine Enterokinase, His, Expressed in Yeast |
100 U/500 U/5000 U |
|
|
Anti-DYKDDDDK(Flag) MagBeads |
1 mL/5 mL |