Cy5-SE (Cy5 NHS Ester) is a reactive dye for the labeling of amino-groups inpeptides, proteins, andoligonucleotides. This dye requires small amount of organic co-solvent (such as DMF or DMSO) to be used in labeling reaction. This reagent is ideal for very cost-efficient labeling of soluble proteins, as well as all kinds ofpeptidesandoligonucleotides. This reagent also works well in organic solvents for small molecule labeling. Excitation (nm):649, Emission (nm): 670.

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Conjugation of SELP Analogues with Cy5-SE.
1. 1 mL of each 100 μg/mL SELP solution in 100 mM sodium bicarbonate buffer (pH 8.3) is mixed with 10 μL of 1.2 mg/mL Cy5 mono NHS-ester in 10% DMSO and incubates for 2 hours on ice.
2. To quench the reaction, 50 μL of 1 M Tris-HCl (pH 8.0) is added to the reaction solution.
3. Reaction mixtures are loaded onto 1.5 mL Sephadex G-25 columns, and Cy5-conjugated SELPs are eluted by centrifugation for 3 minutes at 1050g^[1].

753.88

Solid

C₃₇H₄₃N₃O₁₀S₂

146368-14-1

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

DMSO : 25 mg/mL(33.16 mM;Need ultrasonic)

H₂O : ≥ 5.88 mg/mL(7.80 mM)

*"≥" means soluble, but saturation unknown.

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℃, 6 months; -20℃, 1 month (sealed storage, away from moisture and light)。-80℃ 储存时，请在 6 个月内使用，-20℃ 储存时，请在 1 个月内使用。

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: ≥ 2.5 mg/mL (3.32 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.32 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 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-CDin saline)

  Solubility: ≥ 2.5 mg/mL (3.32 mM); Clear solution

  此方案可获得 ≥ 2.5 mg/mL (3.32 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 25.0 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。

  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

• 
  Description: CY5-SE can be used to label peptides, proteins, and oligonucleotides. CY5-SE can be used to detect the biodistribution in vivo.
  Method: For synthesis of Cy5-conjugated mesoporous silica nanoparticles (MSNs) and detect biodistribution in vivo.
  1. CY5-SE (2.5 mg) and APTES (200 μL) are dissolved in DMSO (2 mL) and the solution reacted for 24 h in the dark under stirring.
  2. CTAB, Brij58 and TEOS are added and reacted to synthesis of small pore MSN and MSNs. Above reaction (200 μL) solution is added.
  3. The male BALB/c nude mice are given 200 μL of 5 mg/mL Cy5-lf-protocells via intraperitoneal injection, with corresponding dose of MSNs and Cy5-protocells as control.
  4. Then, the mice are anesthetized by isoflurane inhalation and scanned in an IVIS Spectrum CT system (Maestro2, CRI, USA) at various time points.
• 
  Description: CY5-SE can be used to label peptides, proteins, and oligonucleotides. CY5-SE can be used to prepare viral nanoparticle to detect the biodistribution in vivo.
  Method: For the preparation of viral nanoparticle and bioimaging.
  1. CY5-SE is added at increasing coat proteins:dye ratios of 1:1, 1:2, 1:5, 1:10 and 1:20.
  2. For fluorescence-labeled particles, virus-Cy5 conjugates are purified by three rounds of ultracentrifugation (112000 g, 1 h, 4℃) over a 30% (w/v) sucrose cushion.
  3. The pellets are then resuspended in 10 mM KP buffer, pH 7.0.
  4. Virus-Cy5 are administered intravenously at a dose of 10 mg/kg body weight.
  5. Mice are euthanized and organs are collected for ex vivo imaging using the IVIS Spectrum Imaging System (PerkinElmer Ltd) 24 h post injection. Regions of interest (ROI) are analyzed using the IVIS Living Image v4.2 (PerkinElmer Ltd) to determine the fluorescence intensity per area from each organ.