FLAG tag Peptide
(Synonyms: H-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH) 目录号 : GP10150
FLAG tag Peptide 是一种包含肠激酶限制性位点的 8 肽 (Asp-Tyr-lys-Asp-Asp-Asp-ASP-ASP-Lys)。
Cas No.:98849-88-8
Sample solution is provided at 25 µL, 10mM.
FLAG tag Peptide is an 8-peptide (Asp-Tyr-lys-Asp-Asp-Asp-ASP-ASP-Lys) containing an intestinal kinase restriction site. FLAG Tag peptide fusion system is used in immunoaffinity chromatography for protein identification and purification. FLAG tag Peptide label peptides allow elution under non-denatured conditions[1,6]. The FLAG tag Peptide can be fused to either the N- or C-terminus of a given fusion protein.
The FLAG tag Peptide marker peptide fusion system comprises a unique and widely useful technique for protein identification and purification. When covalently attached to a solid support, the anti-Flag M1 antibody can be used for the rapid purification of FLAG tag Peptide fusion proteins in a mild, calcium-dependent affinity chromatography procedure. FLAG tag Peptide fusion proteins are typically purified to homogeneity in a single step, starting from a crude cell homogenate or supernatant, without ever exposing the protein to conditions other than physiological saline at pH 7.2 (with calcium or EDTA). The entire purification process can be carried out within several hours[2-3].
The N-terminal FLAG tag Peptide fusion protein can be purified by agarose affinity gel[5]. A short-cut protein purification procedure, where mechanical cell disintegration and affinity purification by immobilization of the anti-Flag M1 antibody on magnetic glass beads are combined. The modified glass beads were added to yeast cells and mixed on a laboratory vortex mixer. The magnetic glass beads capture the target protein, while the yeast cells are disrupted. By making contact with a stationary magnet, the magnetic glass beads can very efficiently be separated from yeast cell debris. Chelating agents, such as EDTA, were capable of eluting the FLAG tag Peptide fusion proteins from the magnetic glass beads[4].
References:
[1]. Einhauer A, Jungbauer A. The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins. J Biochem Biophys Methods. 2001 Oct 30;49(1-3):455-65. doi: 10.1016/s0165-022x(01)00213-5. PMID: 11694294.
[2]. Chiang CM, Roeder RG. Expression and purification of general transcription factors by FLAG epitope-tagging and peptide elution. Pept Res. 1993 Mar-Apr;6(2):62-4. PMID: 7683509.
[3]. Prickett KS, Amberg DC, Hopp TP. A calcium-dependent antibody for identification and purification of recombinant proteins. Biotechniques. 1989 Jun;7(6):580-9. PMID: 2698650.
[4]. Schuster M, Wasserbauer E, Ortner C, Graumann K, Jungbauer A, Hammerschmid F, Werner G. Short cut of protein purification by integration of cell-disrupture and affinity extraction. Bioseparation. 2000;9(2):59-67. doi: 10.1023/a:1008135913202. PMID: 10892539.
[5]. Hopp TP, Prickett KS, Price VL, et al. A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and Purification. Bio/technology (Nature Publishing Company). 1988;6:1204-1210. DOI: 10.1038/nbt1088-1204.
[6].Li Y. Commonly used tag combinations for tandem affinity purification. Biotechnol Appl Biochem. 2010 Feb 15;55(2):73-83. doi: 10.1042/BA20090273. PMID: 20156193.
FLAG tag Peptide 是一种包含肠激酶限制性位点的 8 肽 (Asp-Tyr-lys-Asp-Asp-Asp-ASP-ASP-Lys)。 FLAG Tag 肽融合系统用于免疫亲和层析，用于蛋白质鉴定和纯化。 FLAG 标签肽标签肽允许在非变性条件下洗脱[1,6]。 FLAG 标签肽可以融合到给定融合蛋白的 N 端或 C 端。
FLAG 标签肽标记肽融合系统包含一种独特且广泛有用的蛋白质鉴定和纯化技术。当共价连接到固体支持物上时，抗 Flag M1 抗体可用于在温和的钙依赖性亲和层析过程中快速纯化 FLAG 标签肽融合蛋白。 FLAG 标签肽融合蛋白通常从粗细胞匀浆或上清液开始一步纯化至均质，除了 pH 7.2 的生理盐水（含钙或 EDTA）外，从未将蛋白质暴露于其他条件。整个纯化过程可在数小时内完成[2-3]。
N-terminal FLAG tag Peptide融合蛋白可以通过琼脂糖亲和凝胶纯化[5]。一种快捷的蛋白质纯化程序，其中结合了机械细胞分解和通过将抗 Flag M1 抗体固定在磁性玻璃珠上进行的亲和纯化。将改性玻璃珠添加到酵母细胞中并在实验室涡旋混合器上混合。磁性玻璃珠捕获目标蛋白，同时破坏酵母细胞。通过与固定磁铁接触，磁性玻璃珠可以非常有效地与酵母细胞碎片分离。 EDTA 等螯合剂能够从磁性玻璃珠上洗脱 FLAG 标签肽融合蛋白[4]。
| Purification of ribonucleoprotein (RNP) complexes composed of specific cellular rna by FLAG peptide coupled antisense oligonucleotide (ASO) pull-down [1]: | |
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Preparation Method |
Preparation of FLAG tag Peptide-Conjugated Antisense Oligonucleotide 1.Mix 50 μL of 20 μM (1,000 pmol/50 μL) ice-cold oligonucleotide with 50 μL of freshly prepared cold 1 M NaIO4. 2.Incubate on ice for 10 min. 3.Add 1 mL of ice-cold 2 % LiClO4/acetone and vortex. 4.Incubate on ice for 10 min. 5.Centrifuge the tube at 20,000 g at 4 ℃ for 10 min. 6.Carefully aspirate the supernatant to waste and add 1 mL of cold acetone. 7.Centrifuge the tube at 20,000 g at 4℃ for 5 min. 8.During the centrifugation, prepare the FLAG-hydrazide solution. 9.Aspirate the supernatant to waste. 10.Dissolve the precipitated oligonucleotide in 12 μL of 0.1 M sodium acetate (pH 5.2). 11.Add 12 μL of 30 mM FLAG-peptide-hydrazide solution. 12.Incubate the tube at room temperature for 30 min. 13.Add 10 μL of 1 M NaCNBH3. 14.Incubate the tube at room temperature for 30 min. 15.Add 60 μL of ultrapure water and 10 μL of 3 M sodium acetate (pH 5.2), then vortex. 16.Add 250 μL of 100 % ethanol and vortex. 17.Incubate the tube at −80℃ for 30 min. 18.Centrifuge the tube at 15,000 g for 10 min. 19.Carefully aspirate the supernatant to waste and add 1 mL of 80 % ethanol. 20.Centrifuge the tube at 15,000 g for 5 min. 21.Carefully aspirate the supernatant to waste. Repeat the wash step. 22.Dissolve the oligonucleotide pellet in 50 μL of water. 23.Estimate the concentration of the FLAG-peptide-conjugated-oligonucleotide by measuring the absorbance at 260 nm and comparing with the absorbance of the original oligonucleotide solution. |
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References: [1]. Adachi S, Natsume T. Purification of noncoding RNA and bound proteins using FLAG peptide-conjugated antisense-oligonucleotides. Methods Mol Biol. 2015;1262:265-74. doi: 10.1007/978-1-4939-2253-6_16. PMID: 25555587. |
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| Cas No. | 98849-88-8 | SDF | |
| 别名 | H-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-OH | ||
| 化学名 | FLAG Peptide | ||
| Canonical SMILES | C1=CC(=CC=C1CC(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)N)O | ||
| 分子式 | C41H60N10O20 | 分子量 | 1012.97 |
| 溶解度 | ≥ 50.6 mg/mL in DMSO, ≥ 210.6 mg/mL in Water, ≥ 34.03 mg/mL in EtOH | 储存条件 | Store at -20°C, protect from light, stored under nitrogen |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 0.9872 mL | 4.936 mL | 9.872 mL |
| 5 mM | 0.1974 mL | 0.9872 mL | 1.9744 mL |
| 10 mM | 0.0987 mL | 0.4936 mL | 0.9872 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet