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3Xflag能够使可免疫原性和与抗体的亲和性得到增强。3×FLAG Peptide是一个合成的含有三个重复DYKXXD氨基酸序列的多肽,常用于竞争性结合Anti-Flag抗体。
编号:184546
CAS号:402750-12-3
单字母:H2N-MDYKDHDGDYKDHDIDYKDDDDK-OH
3X FLAG peptide TFA 是一种合成的多肽,包含三个重复的 Asp-Tyr-Lys-Xaa-Xaa-Asp 基序。
Flag多肽为编码8个氨基酸的亲水性多肽(DYKDDDDK),可用于检测特异性anti-Flag单抗,为提高检测灵敏性,现研发出3×Flag重复多肽(3×Flag-tag Peptide),该3×Flag重复多肽含23个氨基酸,可以检测高达10fmol融合蛋白。3×Flag重复多肽已广泛用于纯化Flag融合蛋白,竞争性结合Anti-Flag单抗。
3× FLAG标签系统大幅度提高了FLAG标签的检测能力。三串联的FLAG表位是亲水性的,22个氨基酸的长链,它最多可以检测10 fmol的融合蛋白。FLAG标记的Pyrococcus furiosus的麦芽糊精结合蛋白已经获得了晶体,晶体结果显示FLAG标签对蛋白晶体质量没有多大影响。
FLAG标签可以用肠激酶法处理去除,肠激酶能够特异性的作用于C端的5个氨基酸多肽序列。
用途:3-Flag peptide被广泛用作一种温和的纯化试剂,用于Flag表位标记重组蛋白的纯化。尽管在缺乏钙离子时,其亲和柱会释放单价标记蛋白,但是抗体仍保留对Flag序列的亲和性,即使在无金属存在的条件下,因此不能用来进行金属敏感的ELISA实验。当Flag标记蛋白结合在ELISA板或印迹过滤器上时,抗体仍然与多价表面包被抗原结合。抗原多价性提高了Flag抗体的亲和性,使得ELISA反应不依赖于钙离子。然而,当抗体本身是单价的,即通过蛋白水解变成Fab,ELISA反应即变为钙依赖的。这种金属依赖的ELISA实验被用于研究抗体对金属的需求。在二价金属中,比钙半径大或比钙半径小的金属其结合均逐渐减少。几种更小的金属,比如镍,充当结合反应的抑制剂。重金属如镉、镧和钐可以与抗体结合。由于对该抗体与重组Flag融合蛋白共结晶的兴趣,其与重金属结合的能力就变得非常有意义。
3×FLAG Peptide是一个合成的含有三个重复DYKXXD氨基酸序列的多肽,常用于竞争性结合Anti-Flag抗体。3× Flag Peptide 主要用于蛋白鉴定与纯化,比如可以在免疫沉淀时洗脱与Anti-Flag抗体结合的Flag融合表达蛋白的多肽[1-5]。
3X FLAG Peptide is a synthetic polypeptide containing three repeated DYKXXD amino acid sequences, often used to competitively bind Anti-Flag antibodies.3X Flag Peptide is mainly used for protein identification and purification, such as elution of Flag fusion expression proteins bound to Anti-Flag antibodies during immunoprecipitation[1-5].
References:
[1]. Hopp, T., Prickett, K., et al. A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and Purification. Nat Biotechnol 6, 1204-1210 (1988). https://doi.org/10.1038/nbt1088-1204
[2]. Einhauer A, Jungbauer A. Affinity of the monoclonal antibody M1 directed against the FLAG peptide. J Chromatogr A. 2001 Jun 29;921(1):25-30. doi: 10.1016/s0021-9673(01)00831-7. PMID: 11461009.
[3].Gao XK, Rao XS, et al. Phase separation of insulin receptor substrate 1 drives the formation of insulin/IGF-1 signalosomes. Cell Discov. 2022 Jun 28;8(1):60. doi: 10.1038/s41421-022-00426-x. PMID: 35764611; PMCID: PMC9240053.
[4].Gao J, Huang G, et al.PROTEIN S-ACYL TRANSFERASE 13/16 modulate disease resistance by S-acylation of the nucleotide binding, leucine-rich repeat protein R5L1 in Arabidopsis. J Integr Plant Biol. 2022 Sep;64(9):1789-1802. doi: 10.1111/jipb.13324. Epub 2022 Jul 28. PMID: 35778928.
[5]. Lian H, Jiang K, et al.The Salmonella effector protein SopD targets Rab8 to positively and negatively modulate the inflammatory response. Nat Microbiol. 2021 May;6(5):658-671. doi: 10.1038/s41564-021-00866-3. Epub 2021 Feb 18. PMID: 33603205; PMCID: PMC8085087.
专肽生物可以提供常见抗体制备、免疫学研究等生物科研需求的各类标签多肽(Tag peptide)以及蛋白底物产品,可用荧光标记(FITC、5-Fam)等。除常见抗体中抗原决定部位的标签多肽(tag: His | c-Myc | HA | FLAG | V5 | X-press | VSV tag | T7 tag | Snoop tag | E-tag | Sof tag 1 | Spy tag |等等),我们还提供各类定制标签肽的合成,并可提供荧光标记(FITC、5-Fam)等。同时,免费提供的MS和HPLC分析报告。
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