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

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

 Improved Yield and Purity

Figure 1. Anti-DYKDDDDK (Flag) Affinity Gel has higher yield and purity of secretory or membrane proteins compared to other suppliers.

Publication

1. 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)
2. 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)
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)
4. 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)
5. 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)
6. 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)
7. 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)
8. 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)
9. 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)
10. 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

20585_MSDS_HB250806_EN.PDF

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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