Biotin NHS是用于制备生物素化表面或多肽的氨基反应性生物素试剂。
生物活性 | Biotin NHS is an amino reactive biotin reagent used in the preparation of biotinylated surfaces or polypeptides. |
体外研究 (In Vitro) | Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs). Calculations: For best results, use ≥ 12-fold molar excess of biotin for a 10 mg/mL protein solution or ≥ 20-fold molar excess of biotin for a 2 mg/mL protein solution. 1.Calculate millimoles of biotin reagent to add to the reaction for a 20-fold molar excess: mL protein × (mg protein/mL protein) × (mmoL protein/mg protein) × (20 mmoL protein/mmoL protein) = mmoL Biotin. * 20 = Recommended molar fold excess of biotin for 2 mg/ml protein sample 2. Calculate microliters of 10 mM biotin reagent solution (prepared in Step B.3) to add to the reaction: mmoL Biotin × (1000000 μL/L) × (L/ 10 mmoL) =μL Biotin Biotin Labeling Reaction: 1. Dissolve 1-10 mg protein in 0.5-2.0 mL PBS according to the calculation made in Calculations. 2. Before use, prepare a 10 mM solution of the biotin reagent in an organic solvent such as DMSO. 3. Add the appropriate volume (μL) of 10 mM biotin reagent solution to the protein solution. 4. Incubate reaction on ice for two hours or at room temperature for 30 minutes. 5. To tests the labeled protein by ELISA or Western blot.
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运输条件 | Room temperature in continental US; may vary elsewhere. |
储存方式 | -20°C, sealed storage, away from moisture and light *该产品在溶液状态不稳定,建议您现用现配,即刻使用。 |
溶解性数据 | In Vitro: DMSO : 62.5 mg/mL(183.08 mM;Need ultrasonic) 配制储备液 1 mM | 2.9293 mL | 14.6464 mL | 29.2929 mL | 5 mM | 0.5859 mL | 2.9293 mL | 5.8586 mL | 10 mM | 0.2929 mL | 1.4646 mL | 2.9293 mL |
*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;该产品在溶液状态不稳定,建议您现用现配,即刻使用。 In Vivo: 请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照In Vitro方式配制澄清的储备液,再依次添加助溶剂: ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用;
以下溶剂前显示的百 分比是指该溶剂在您配制终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶 1. 请依序添加每种溶剂: 10% DMSO 40%PEG300 5%Tween-80 45% saline Solubility: ≥ 2.08 mg/mL (6.09 mM); Clear solution
此方案可获得 ≥ 2.08 mg/mL (6.09 mM,饱和度未知) 的澄清溶液。 以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中,混合均匀;向上述体系中加入50 μL Tween-80,混合均匀;然后继续加入 450 μL生理盐水定容至 1 mL。 2. 请依序添加每种溶剂: 10% DMSO 90% (20%SBE-β-CDin saline) Solubility: ≥ 2.08 mg/mL (6.09 mM); Clear solution
此方案可获得 ≥ 2.08 mg/mL (6.09 mM,饱和度未知) 的澄清溶液。 以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL20% 的 SBE-β-CD 生理盐水水溶液中,混合均匀。 3. 请依序添加每种溶剂: 10% DMSO 90%corn oil Solubility: ≥ 2.08 mg/mL (6.09 mM); Clear solution
此方案可获得 ≥ 2.08 mg/mL (6.09 mM,饱和度未知) 的澄清溶液,此方案不适用于实验周期在半个月以上的实验。 以 1 mL 工作液为例,取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中,混合均匀。 *以上所有助溶剂都可在本网站选购。
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染色示例 | Description: Biotin NHS is an amino reactive biotin reagent, it can be used as a chemical label for polypeptide and nanocarrier.Method: For chemical labeling use.1. A 1:1 molar ratio of Biotin NHS is added to the ligand of nanocarrier (0.5 mg/mL).2. The reaction is allowed to proceed for two hours in PBS, pH 7.4 at room temperature.3. Free biotin is removed using a 10-kDa cutoff spin filters (10 washes with PBS).4. Use a transmission electron microscopy for image. Description: Biotin NHS is an amino reactive biotin reagent, it can be used to label biotinylated virus.Method: For virus labeling.1. Biotin NHS is added at a 1:1 molar ratio with the coat proteins.2. Biotinylated viruses are purified by 10 washes in 10 mM KP buffer, and use 100-kDa molecular weight cutoff size exclusion spin filters.3. Use a transmission electron microscopy for image. Description: Biotin NHS is an amino reactive biotin reagent, it can be used to label biotinylated virus.Method: For viruses labeling.1. Viruses is used at the final concentration of 2 mg/mL in 0.1 M potassium phosphate buffer (KP buffer, pH 7.0) and mixed with Biotin NHS (in DMSO) for reactions. The final concentration of DMSO in the reaction solution was adjusted to 10% (v/v).2. Mixtures are agitated at room temperature overnight.3. Particles are purified using 10-kDa molecular weight cutoff size exclusion spin columns.4. Use a transmission electron microscopy for image.
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