| Field | Specification |
|---|---|
| Mfr No | |
| Form | suspension (agarose slurry) |
| Product Type | |
| Shipping | |
| Storage |
Description
Anti-Flag affinity purification gel (secretory protein or Membrane Protein) is prepared by covalent coupling of high-quality mouse IgG2b monoclonal antibody (clone number: 3F5) and 4% highly cross-linked agarose gel. This product has a high loading capacity of Flag-tagged fusion protein (at least 1.1 mg protein/mL gel) and less non-specific binding of impurities. It can be used for purification and immunoprecipitation (IP) of fusion proteins (secretory protein or membrane protein) with Flag tags.
Anti-Flag affinity purification gel (secretory protein or Membrane Protein) can bind to Met-modified N-terminal Flag fusion protein (Met-FLAG–Protein), N-terminal Flag fusion protein (FLAG–Protein), and C-terminal Flag fusion protein (Protein-FLAG).
Features
- High specificity: High-quality mouse IgG2b monoclonal antibody is used, which can specifically adsorb the Flag protein sequence.
- High loading capacity: Loading capacity ≥ 1.1 mg protein/mL gel.
- High purity: Less non-specific binding of impurities, resulting in high-purity Flag-tagged protein.
- Wide range of uses: Can be used for protein purification or IP.
- Multiple types of binding proteins: Can bind to Met-modified N-terminal Flag fusion protein (Met-Flag–Protein), N-terminal Flag fusion protein (Flag–Protein), C-terminal Flag fusion protein (Protein-Flag)
Specifications
|
Clone |
3F5 |
|
Isotype |
Mouse IgG2b |
|
Application |
Purification of secreted or membrane proteins, immunoprecipitation（IP） |
|
Protein |
≥1.1 mg protein/mL gel |
|
Buffer |
TBS ，50%glycerin ，pH7.4 ，contains 0.02% (w/v) sodium azide |
Shipping and Storage
This product should be stored at 2~8℃ for 1 year. Cryopreservation in solutions without glycerol is prohibited!
Application
Purification of secreted or membrane proteins, immunoprecipitation（IP
Figures
1. High specificity and efficiently
Figure 1. Exhibits high specificity and efficiently binds Flag tags at any position, even at very low target protein expression levels using Anti-DYKDDDDK (Flag) Affinity Gel
2. Improved target protein yield and purity
Figure 2. Anti-DYKDDDDK (Flag) Affinity Gel has higher yield and purity of secretory or membrane proteins compared to other suppliers.
Publication
- Xiao J, Wang S, Chen L, et al. 25-Hydroxycholesterol regulates lysosome AMP kinase activation and metabolic reprogramming to educate immunosuppressive macrophages. Immunity. 2024;57(5):1087-1104.e7. doi:10.1016/j.immuni.2024.03.021(IF:32.4)
- Zheng Z, Zeng X, Zhu Y, et al. CircPPAP2B controls metastasis of clear cell renal cell carcinoma via HNRNPC-dependent alternative splicing and targeting the miR-182-5p/CYP1B1 axis. Mol Cancer. 2024;23(1):4. Published 2024 Jan 6. doi:10.1186/s12943-023-01912-w(IF:37.3)
- Lin J, Jiang X, Dong M, et al. Hepatokine Pregnancy Zone Protein Governs the Diet-Induced Thermogenesis Through Activating Brown Adipose Tissue. Adv Sci (Weinh). 2021;8(21):e2101991. doi:10.1002/advs.202101991(IF:16.806)
- Han L, Zhang F, Liu Y, et al. Uterus globulin associated protein 1 (UGRP1) binds podoplanin (PDPN) to promote a novel inflammation pathway during Streptococcus pneumoniae infection. Clin Transl Med. 2022;12(6):e850. doi:10.1002/ctm2.850(IF:11.492)
- Chen X, Duan Y, Qiao F, et al. A secreted fungal effector suppresses rice immunity through host histone hypoacetylation [published online ahead of print, 2022 May 20]. New Phytol. 2022;10.1111/nph.18265. doi:10.1111/nph.18265(IF:10.152)
- Chen Y, Yang L, Lu Y, et al. Up-regulation of NMRK2 mediated by TFE3 fusions is the key for energy metabolism adaption of Xp11.2 translocation renal cell carcinoma. Cancer Lett. 2022;538:215689. doi:10.1016/j.canlet.2022.215689(IF:8.679)
- Chen X, Li X, Li P, et al. Comprehensive identification of lysine 2-hydroxyisobutyrylated proteins in Ustilaginoidea virens reveals the involvement of lysine 2-hydroxyisobutyrylation in fungal virulence. J Integr Plant Biol. 2021;63(2):409-425. doi:10.1111/jipb.13066(IF:7.061)
- Tang C, Zhou Y, Sun W, et al. Oncopeptide MBOP Encoded by LINC01234 Promotes Colorectal Cancer through MAPK Signaling Pathway. Cancers (Basel). 2022;14(9):2338. Published 2022 May 9. doi:10.3390/cancers14092338(IF:6.639)
- Zheng G, Jiang C, Li Y, et al. TMEM43-S358L mutation enhances NF-κB-TGFβ signal cascade in arrhythmogenic right ventricular dysplasia/cardiomyopathy. Protein Cell. 2019;10(2):104-119. doi:10.1007/s13238-018-0563-2(IF:6.228)
- Chen X, Tang J, Pei Z, et al. The 'pears and lemons' protein UvPal1 regulates development and virulence of Ustilaginoidea virens. Environ Microbiol. 2020;22(12):5414-5432. doi:10.1111/1462-2920.15284(IF:4.933)
FAQ
Q: If the protein is to remain active, which substances should be used for elution?
A: Non-denaturing elution method can be used.
Q: How does this product compare with those from the imported manufacturers?
A: The protein binding capacity of Yeasen 20585 is nearly twice that of the imported S* brand.
Q: We used the flag peptide for elution, but it couldn't be completely removed?
A: The loading capacity of the medium is already relatively low, and the elution concentration might be quite low. There is a possibility that due to the low concentration, no distinct bands will be visible. It is recommended to increase the elution intensity in the next step.
Documents:
Safety Data Sheet
Manuals
20585_Manual_Ver.EN20250806.pdf
Related Blog:
Anti-Flag Affinity Gel: A Reliable Partner for IP and Protein Purification
Yeasen Anti-DYKDDDDK (Flag) Affinity Gel: Precision Purification for Low-Abundance Membrane Proteins
The Tool for Capturing Flag/HA/Myc-Tagged Proteins
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The key difference is the matrix: MagBeads use superparamagnetic polymer beads enabling magnetic separation (no centrifugation needed, fast processing); Affinity Gel uses 4% highly cross-linked agarose (Sepharose-type) separated by gravity flow or low-speed centrifugation. The Affinity Gel (clone 3F5) is particularly designed for secretory and membrane proteins and provides a higher binding capacity (≥1.1 mg/mL gel).
Yes — the Anti-Flag Affinity Gel (clone 3F5) is specifically noted for purification of secretory and membrane proteins, in addition to intracellular proteins. The agarose matrix is compatible with mild detergent conditions (e.g., DDM, digitonin) commonly used to solubilize membrane proteins, though detergent compatibility should be validated for your specific conditions.
Preferred: competitive elution with Flag peptide (DYKDDDDK) at 100–500 µg/mL in wash buffer — provides native, functional protein. Alternative: low pH (glycine, pH 3.0–3.5) for non-peptide elution; neutralize immediately after collection. Flag peptide elution is gentler and better preserves protein activity.
Yes — agarose-based affinity gels can typically be regenerated for multiple uses. After elution, wash with 5–10 column volumes of low-pH buffer, then re-equilibrate with TBS. Regeneration efficiency decreases over repeated use. The gel should be stored in TBS with 50% glycerol and 0.02% sodium azide at 2–8°C between uses.
Store at 2–8°C for 1 year. Do not freeze without glycerol — cryopreservation in solutions without glycerol is prohibited. Storage buffer: TBS, 50% glycerol, pH 7.4, 0.02% sodium azide.
The following customization and add-on services may be available for this product type through the supplier. For inquiries and pricing, contact support@biohippo.com.
Customization Options
- Bulk Quantities: Larger volumes and custom pack sizes beyond the standard catalog may be available for high-throughput core facilities and industrial customers. Contact us to inquire about bulk pricing.
- Custom Antibody Coupling (Magnetic Beads): Custom magnetic beads conjugated to your specific antibody or protein ligand may be available through Yeasen's OEM and custom services program. These services leverage the same bead platform used for catalog products.
- OEM Manufacturing: Custom bead formulations for diagnostic device development, including magnetic beads for nucleic acid isolation, IP-grade beads, and affinity capture beads, may be available at manufacturing scale.
To inquire about custom options or bulk pricing, contact support@biohippo.com.
