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3X FLAG Peptide

Catalog No.
A6001
合成的标签肽。
组合的产品项目
规格价格库存 数量
4mg
¥ 900.00
现货
25mg
¥ 4,050.00
现货

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Background

The FLAG-tag system utilizes a short, hydrophilic 8- amino-acid peptide that is fused to the protein of interest1. The FLAG peptide binds to the antibody M1. Whether binding is calcium-dependent manner2 or –independent3 remains controversial. A disadvantage of the system is that the monoclonalantibody purification matrix is not as stable as others. In general, small tags can be detected with specific monoclonal antibodies.

To improve the detection of the FLAG tag the 3x FLAG system has been developed. This threetandem FLAG epitope is hydrophilic, 22-amino-acids long and can detect up to 10 fmol of expressed fusion protein. The FLAG-tagged maltodextrin-binding protein of Pyrococcus furiosus has been crystallized4 and the quality of the crystals was very similar to that of crystals of untagged protein.

 Finally, the FLAG-tag can be removed by treatment with enterokinase, which is specific for the five C-terminal amino acids of the peptide sequence5.

References:
1. Hopp TP, Prickett KS, Price VL, Libby RT, March CJ, Ceretti DP, Urdal DL, Conlon PJ (1988) A short polypeptide marker sequence useful for recombinant protein identification and purification. Bio/Technology 6:1204–1210.
2. Hopp TP, Gallis B, Prikett KS (1996) Metal-binding properties of a calcium dependent monoclonal antibody. Mol Immunol 33:601–608.
3. Einhauer A, Jungbauer A (2000) Kinetics and thermodynamical properties of the monoclonal antibody M1 directed against the FLAG peptide. 20th International symposium on the separation of proteins, peptides, and polynucleotides (ISPPP). Lublijana, Slovenia, November 5–8, 2000.
4. Bucher MH, Evdokimov AG, Waugh DS (2002) Differential effects of short affinity tags on the crystallization of Pyrococcus furiosus maltodextrin-binding protein. Biol Cryst 58:392–397.
5. Maroux S, Baratti J, Desnuelle P (1971) Purification and specificity of procine enterokinase. J Biol Chem 246:5031–5039.

文献引用

Chemical Properties

StorageDesiccate at -20°C
M.Wt2861.87
FormulaC120H169N31O49S
SynonymsH-Met-Asp-Tyr-Lys-Asp-His-Asp-Gly-Asp-Tyr-Lys-Asp-His-Asp-Ile-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH
Solubility≥25mg/ml in TBS (0.5M Tris-HCl, pH 7.4, with 1M NaCl), 1X TBS as recommended buffer.
Chemical Name3X FLAG Peptide
SDFDownload SDF
Canonical SMILESCCC(C)C(C(=O)NC(CC(=O)O)C(=O)NC(CC1=CC=C(C=C1)O)C(=O)NC(CCCCN)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CC(=O)O)C(=O)NC(CCCCN)C(=O)O)NC(=O)C(CC(=O)O)NC(=O)C(CC2=CNC=N2)NC(=O)C(CC(=O)O)NC(=O)C(CCCCN)NC(=O)C(CC3=CC=C(C=C3)O)NC(=O)C(CC(=O)O)NC(
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

ELISA实验[1]:

溶解方法

该多肽在无菌水中的溶解度>10 mM,原液应分装并储存在-80℃,可储存几个月。

应用

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。

References:

1. Hopp TP1, Gallis B, Prickett KS. Metal-binding properties of a calcium-dependent monoclonal antibody. Mol Immunol. 1996 May-Jun;33(7-8):601-8.

生物活性

描述 3X FLAG Peptide是一个合成的含有三个重复DYKXXD氨基酸序列的多肽。
靶点 anti-flag M2 antibody          
IC50            

质量控制

质量控制和MSDS

批次:

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