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- 详细信息
- 文献和实验
- 技术资料
- 免疫原:
Recombinant DYKDDDDK tag protein (Contains same sequence as FLAG® Tag)
- 亚型:
IgG
- 保存条件:
Store at -20°C. Avoid exposure to light. Stable for one year after shipment.
- 克隆性:
Polyclonal
- 标记物:
CoraLite® Plus 750 Fluorescent Dye
- 适应物种:
recombinant protein
- 宿主:
Rabbit
- 应用范围:
WB, FC (Intra)
- 靶点:
DYKDDDDK tag
- 抗体英文名:
CoraLite® Plus 750-conjugated DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope)
- 抗体名:
CoraLite® Plus 750-conjugated DYKDDDDK tag Polyclonal antibody (Binds to FLAG® tag epitope)
- 规格:
100ul
经过测试的应用
| Positive WB detected in | Transfected HEK-293 cells |
| Positive FC (Intra) detected in | Transfected HEK-293 cells |
推荐稀释比
| 应用 | 推荐稀释比 |
|---|---|
| Western Blot (WB) | WB : 1:2000-1:16000 |
| Flow Cytometry (FC) (INTRA) | FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension |
| It is recommended that this reagent should be titrated in each testing system to obtain optimal results. | |
| Sample-dependent, Check data in validation data gallery. | |
产品信息
CL750-20543 targets DYKDDDDK tag in WB, FC (Intra) applications and shows reactivity with recombinant protein samples.
| 经测试应用 | WB, FC (Intra) |
| 经测试反应性 | recombinant protein |
| 免疫原 |
CatNo: Ag2329 Product name: Recombinant DYKDDDDK tag protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: flag of Sequence: DYKDDDDK |
| 宿主/亚型 | Rabbit / IgG |
| 抗体类别 | Polyclonal |
| 产品类型 | Antibody |
| 全称 | DYKDDDDK tag |
| 别名 | Flag, DYKDDDDK, DDDDK tag, DDDDK |
| 基因名称 | |
| Gene ID (NCBI) | |
| RRID | AB_3673693 |
| 偶联类型 | CoraLite® Plus 750 Fluorescent Dye |
| 最大激发/发射波长 | 755 nm / 780 nm |
| 形式 | Liquid |
| 纯化方式 | Antigen affinity purification |
| UNIPROT ID | FLAGTAG |
| 储存缓冲液 | PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| 储存条件 | Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. |
背景介绍
Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. The DYKDDDDK(FLAG) peptide has been used extensively as a general tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion. N-terminal DDDDK vectors provide an Ek cleavage site for removal of the fusion tag. The DDDDK peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the DDDDK peptide is more likely to be accessible to antibodies. A DDDDK-tag can be used in many different assays that require recognition by an antibody, such as western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization and so on. This antibody is a rabbit polyclonal antibody raised against 3xFlag (3xDYKDDDDKT) sequence and recognizes the (1x) and (3x)DYKDDDDK peptide and detects DDDDK-tagged proteins. Anti-FLAG is a registered trademark of Sigma-Aldrich Biotechnology.
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文献和实验,在中性和弱碱性条件下可以和固定化的金属离子相互作用(如Ni离子,Zn离子,Co离子等),从而达到纯化蛋白的目的。此系统的洗脱方式可以用过三种非特异性步骤达成: 降低pH(4.5-6) b. 加入EDTA C.不同浓度咪唑溶液(20-250mM) 当蛋白洗脱完毕后,使用EDTA使纯化树脂与金属离子分离,即可完成树脂的再生。 Strep-tag®则是基于链霉亲和素(Streptavidin)和生物素这一自然的相互作用,将Strep-tag®II(WSHPQFEK)或Twin-Strep-tag®
-His-Pro-Gln-Phe-Glu-Lys) 可放在N- 或 C-terminus 不影响蛋白折叠和功能,无需移除 分子量 1058Da Twin-Strep-tag® (也有叫做双Strep-tag®II) 28 aa 标签(WSHPQFEK-GGGSGGGSGG-SA-WSHPQFEK) 两段Strep-tag®II标签序列 较Strep-tag®II对于Strep-Tactin®有更高亲和力 分子量 2887Da 二者均为小标签,不会对蛋白质的结构和功能活性产生影响,因此通常不需要在纯化
IBA 第三代 Strep-tag® 高效蛋白纯化系统,由 Twin-Strep-tag® 搭配全新研发出的 Strep-Tactin®XT 组合而成。在此系统中,Strep-Tactin®XT 与 Twin-Strep-tag® 的亲和力提升至 pM 的范围,与 Strep-tag®II 的亲和力也已升至 nM 的范围。 亲和力大幅改善,其间的链结仍保有可逆性,纯化过程同样温和。仅需简单直接的再生步骤,填料即可重复利用。Strep-Tactin®XT 提升的亲和力能确保您在温和的生理纯化条件
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