This page describes a two-vector retrovirus format for inducible expression.Click this link to learn more about our all-in-one Retro-X Tet-One inducible expression systems.
The Retro-X Tet-On 3G Inducible Expression System combines the efficiency of retroviral gene transfer with the power of our premium tetracycline-regulated gene expression technology.
This page describes a two-vector retrovirus format for inducible expression. Click this link to learn more about our all-in-one Retro-X Tet-One inducible expression systems.
The Retro-X Tet-On 3G Inducible Expression System combines the efficiency of retroviral gene transfer with the power of our premium tetracycline-regulated gene expression technology. The retroviral vectors in this system can be used to establish a tightly controlled inducible gene expression system in a wide variety of dividing hard-to-transfect cell types.
How it works
Doxycycline-controlled transactivators are fusion proteins that contain a DNA-binding TetR domain joined to three minimal transcription activation domains from VP16 (Tet-On 3G, Tet-On Advanced and Tet-Off Advanced proteins). With very high specificity, these transactivators recognize and bind to a tetracycline response element (TRE) that is located in the PTRE3GV promoter upstream of your gene of interest. Binding of the transactivator to the TRE promoter results in strong activation of the downstream gene. The Tet-On 3G system is the premium, third-generation version of the technology, with the tightest TRE promoter and most sensitive transactivator.
In Tet-On systems, gene expression is activated by the addition of Dox to the growth medium.
Ideal retroviral packaging
Retro-X Tet-On 3G systems are supplied with our Retro-X Universal Packaging System (Cat. No. 631530) which produces high titers of MMLV retrovirus and allows you to customize the tropism of your retrovirus to efficiently transduce any cell line or primary cell culture, and thus greatly expands the applications for these inducible systems.
Retro-X Q vector technology
The vectors in these systems feature Takara Bio"s Retro-X Q-Series technology, which:
- Drives high-titer virus production in packaging cells
- Inactivates viral LTRs in target cells to prevent promoter interference
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你用BSA封闭没什么意思，可以换一种，比如明胶，酪蛋白等。
将HRP标记BSA稀释成一定的梯度。
（2）6 X His tag 序列：CAT CAT CAC CAT CAC CAT
（3）GST标签序列：
1ATGTCCCCTATACTAGGTTATTGGAAAATTAAGGGCCTTGTGC
301 AACCCACTCGACTTCTTTTGGAATATCTTGAAGAAAAATATGAAGAGCAT
351 TTGTATGAGCGCGATGAAGGTGATAAATGGCGAAACAAAAAGTTTGAATT
401 GGGTTTGGAGTTTCCCAATCTTCCTTATTATATTGATGGTGATGTTAAAT
451 TAACACAGTCTATGGCCATCATACGTTATATAGCTGACAAGCACAACATG
501 TTGGGTGGTTGTCCAAAAGAGCGTGCAGAGATTTCAATGCTTGAAGGAGC
551 GGTTTTGGATATTAGATACGGTGTTTCGAGAATTGCATATAGTAAAGACT
601 TTGAAACTCTCAAAGTTGATTTTCTTAGCAAGCTACCTGAAATGCTGAAA
651 ATGTTCGAAGATCGTTTATGTCATAAAACATATTTAAATGGTGATCATGT
701 AACCCATCCTGACTTCATGTTGTATGACGCTCTTGATGTTGTTTTATACA
751 TGGACCCAATGTGCCTGGATGCGTTCCCAAAATTAGTTTGTTTTAAAAAA
801 CGTATTGAAGCTATCCCACAAATTGATAAGTACTTGAAATCCAGCAAGTA
851 TATAGCATGGCCTTTGCAGGGCTGGCAAGCCACGTTTGGTGGTGGCGACC
901 ATCCTCCAAAATCGGATCTGGTTCCGCGTGGATCCCCGGAATTCCCGGGT
951 CGACTCGAGCGGCCGCATCGTGACTGA
百度上的，，，，，搬运
Flag标签系统是已被公认的用于表达、纯化以及检测融合蛋白的表达系统，广泛应用于Westernblotting、免疫细胞组化、免疫共沉淀、流式细胞术、蛋白纯化以及蛋白相互作用、蛋白分离等多个研究领域。Flag标签是一个与重组蛋白相融合的由8个亲水氨基酸（Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys）组成的多肽片断。FLAG标签只有8个氨基酸组成，因此它不会占据其他表位与结合域，避免导致融合蛋白的功能、分泌性以及转运发生改变。由于它的亲水性特点，FLAG标签倾向于定位在融合蛋白表面，因此更易与抗体结合以及被肠激酶酶解。
由于Flag标签可位于蛋白质的C端或N端，这一系统已用于各种细胞类型，包括细菌、酵母和哺乳细胞。由于Flag标签系统的纯化条件是非变性的，因此可以纯化所有有活性的融合蛋白。EarthOx的Anti-FlagTagMonoclonalAntibody可以用于检测和Flag标签融合表达蛋白的表达、细胞内定位，以及纯化、定性或定量检测Flag融合表达蛋白等。比如在免疫荧光IF的实验里，同时配合EarthOx的Dylight549荧光二抗，使用该抗体可以准确的定位出Flag融合表达蛋白在细胞内的位置（如图红色显示部分）。另外，做融合表达分析前，通过免疫共沉淀技术获得Flag融合表达蛋白也是非常重要的一步，因此选择一个合适的、可用于免疫共沉淀（Immunoprecipitation，IP）Flag标签抗体也是很必要的。EarthOx的Anti-FlagTagMonoclonalAntibody不仅能应用于细胞内定位的IF实验，同样还能应用于IP实验。1:200甚至更高的IP实验稀释比率，也说明了该抗体本身的具有很高的效价，同时，因为是单克隆抗体，因此特异性也很高。高效价所对应的高稀释比率也意味着超高性价比。
抗体指机体的免疫系统在抗原刺激下,由B淋巴细胞分化成的浆细胞所产生的、可与相应抗原发生特异性结合反应的免疫球蛋白.
载体,你说的是那种载体?生命科学载体有2种,一种是分布于膜上的载体蛋白,和物质运输有关.另一种是基因工程里面的携带外源DNA的一段DNA片段,例如质粒、噬菌体和动植物病毒.
所以,抗体全部是蛋白质,载体部分是蛋白质.
