Biotin NHS is an amino reactive biotin reagent used in the preparation of biotinylated surfaces or polypeptides.

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.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

341.38

C₁₄H₁₉N₃O₅S

35013-72-0

生物素N-羟基琥珀酰亚胺酯

Room temperature in continental US; may vary elsewhere.

-20deg;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。

In Vitro:;

DMSO : 62.5 mg/mL (183.08 mM; Need ultrasonic)

*

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。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in 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 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 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。

• [1]. Wojchowski DM et al. Hybridoma production by simplified avidin-mediated electrofusion. J Immunol Methods. 1986 Jun 24;90(2):173-7.

  [2]. Ngo TT et al. Biotinyl-glucose-6-phosphate dehydrogenase preparation, kinetics, and modulation by avidin. Appl Biochem Biotechnol. 1982 Nov;7(6):443-54.