Biotin-tyramide
(Synonyms: 生物素基酪氨酰胺) 目录号 : GC13376
A reagent used for tyramide signal amplification
Cas No.:41994-02-9
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Tissue |
Brain tissue from Long–Evans rats |
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Preparation Method |
Brain tissue from six animals was processed in one run to fluorescently co-label ARs and NeuN, with and without Biotin-tyramide, as well as ARs and GFAP, with and without Biotin-tyramide. |
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Reaction Conditions |
Biotin-tyramide for 30min |
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Applications |
Biotin-tyramide signal amplification (TSA) dramatically increases AR immunoreactivity in the rat brain, including critical regions of the PFC such as the medial PFC (mPFC) and orbitofrontal cortex (OFC). Biotin-tyramide signal amplification is useful for AR detection with both chromogenic and immunofluorescent immunohistochemistry. |
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References: [1]. Low KL, Ma C, et,al. Tyramide Signal Amplification Permits Immunohistochemical Analyses of Androgen Receptors in the Rat Prefrontal Cortex. J Histochem Cytochem. 2017 May;65(5):295-308. doi: 10.1369/0022155417694870. Epub 2017 Mar 1. PMID: 28438093; PMCID: PMC5407533. |
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Biotin-tyramide is used for tyramide signal amplification (TSA) which is a powerful, patented technology that significantly enhances both chromogenic and fluorescent signals. TSA is easily integrated into standard nonradioactive in situ hybridization (ISH) or IHC protocols.
TSA is an enzyme-mediated detection method that uses horseradish peroxidase (HRP) to catalyze the deposition of a fluorophore-labeled tyramide amplification reagent onto tissue sections or cell preparation surfaces that have been previously blocked with proteins.
Biotin-tyramide use horseradish peroxidase (HRP) to catalyze covalent deposition of biotin labels directly adjacent to the immobilized enzyme. The labeling reaction is quick (less than 10 minutes) and deposited labels can be detected with streptavidin conjugates for imaging in brightfield or fluorescence microscopy.
生物素-酪胺用于酪胺信号放大 (TSA)，这是一种强大的专利技术，可显着增强显色和荧光信号。 TSA 很容易集成到标准的非放射性原位杂交 (ISH) 或 IHC 方案中。
TSA 是一种酶介导的检测方法，它使用辣根过氧化物酶 (HRP) 催化荧光团标记的酪胺扩增试剂沉积到先前已用蛋白质封闭的组织切片或细胞制备表面。
生物素-酪胺使用辣根过氧化物酶 (HRP) 催化直接与固定化酶相邻的生物素标记共价沉积。标记反应很快（不到 10 分钟），沉积的标记可以用链霉亲和素偶联物检测，以便在明场或荧光显微镜下成像。
| Cas No. | 41994-02-9 | SDF | |
| 别名 | 生物素基酪氨酰胺 | ||
| 化学名 | N-(4-hydroxyphenethyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide | ||
| Canonical SMILES | O=C1N[C@@]([H])([C@]([H])(CS2)N1)[C@@H]2CCCCC(NCCC3=CC=C(O)C=C3)=O | ||
| 分子式 | C18H25N3O3S | 分子量 | 363.47 |
| 溶解度 | ≥ 72.6mg/mL in DMSO, ≥ 8.18 mg/mL in EtOH with ultrasonic | 储存条件 | Store at -20°C |
| 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 | 2.7513 mL | 13.7563 mL | 27.5126 mL |
| 5 mM | 0.5503 mL | 2.7513 mL | 5.5025 mL |
| 10 mM | 0.2751 mL | 1.3756 mL | 2.7513 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。
APEX Peroxidase-Catalyzed Proximity Labeling and Multiplexed Quantitative Proteomics
Methods Mol Biol2019;2008:41-55.PMID: 31124087DOI: 10.1007/978-1-4939-9537-0_4
Peroxidase-catalyzed proximity labeling is a powerful technique for defining the molecular environment of proteins in vivo. Expressing a protein of interest fused to a modified plant peroxidase (APEX2) allows labeling of nearby polypeptides. Addition of hydrogen peroxide (H2O2) and biotin-tyramide (biotin-phenol) generates short-lived radicals around the peroxidase. Labeling is thus restricted to proteins in close proximity, providing a snapshot of the local environment around the APEX2 fusion protein. Combined with an initial perturbation, progressive changes in interaction partners can be tracked, e.g., after drug treatment. Multiplexed quantitative mass spectrometry permits the parallel analysis of several experimental replicates or of up to 11 time points. Here we describe the denaturing purification of biotin-labeled proteins with magnetic streptavidin beads, and subsequent sample preparation for multiplexed quantitative mass spectrometry. Proximity-labeled proteins are enriched under strong denaturing conditions. Tryptic on-bead digest of purified proteins is combined with tandem mass tag peptide labeling (TMT), alkaline reversed-phase peptide fractionation, and SPS MS3 mass spectrometry. This analysis pipeline enables studies of complex protein environment changes in perturbed biological systems, as well as comparative studies of functional protein proximity in different cell lines. Through multiplexing, hundreds of proteins can be quantified in each experimental condition in parallel.
Tyramide Signal Amplification Permits Immunohistochemical Analyses of Androgen Receptors in the Rat Prefrontal Cortex
J Histochem Cytochem2017 May;65(5):295-308.PMID: 28438093DOI: 10.1369/0022155417694870
Research on neural androgen receptors (ARs) has traditionally focused on brain regions that regulate reproductive and aggressive behaviors, such as the hypothalamus and amygdala. Although many cells in the prefrontal cortex (PFC) also express ARs, the number of ARs per cell appears to be much lower, and thus, AR immunostaining is often hard to detect and quantify in the PFC. Here, we demonstrate that biotin tyramidesignal amplification (TSA) dramatically increases AR immunoreactivity in the rat brain, including critical regions of the PFC such as the medial PFC (mPFC) and orbitofrontal cortex (OFC). We show that TSA is useful for AR detection with both chromogenic and immunofluorescent immunohistochemistry. Double-labeling studies reveal that AR+ cells in the PFC and hippocampus are NeuN+ but not GFAP+ and thus primarily neuronal. Finally, in gonadally intact rats, more AR+ cells are present in the mPFC and OFC of males than of females. Future studies can use TSA to further examine AR immunoreactivity across ages, sexes, strains, and different procedures (e.g., fixation methods). In light of emerging evidence for the androgen regulation of executive function and working memory, these results may help understand the distribution and roles of ARs in the PFC.