Cyanine7.5 amine is a near-infrared Cy7.5-based dye and also a substrate for amide bond coupling (Ex/Em = 788 nm/808 nm). Cyanine7.5 amine contains a free amino group and can be used for coupling with activated carboxylic acid derivatives [1].
In Vitro, Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.) 1. Preparation of Reagents: (1) Dye Solution: Dissolve Cyanine7.5 amine in dry DMF or DMSO to create a 10-50 mg/mL stock solution. (2) Biomolecule Solution: Dissolve the protein/antibody in a coupling buffer (e.g., 50 mM borate buffer or PBS, pH 8.3-8.5) at a concentration of 1-10 mg/mL. 2. Reaction Procedure: (1) Slowly add the Cyanine7.5 amine stock solution into the protein solution while stirring. Use a molar excess of dye (typically 5-20 fold relative to the protein). (2) Incubate the mixture at room temperature for 1-2 hours or at 4°C overnight, protected from light. 3. Purification: Remove unreacted dye using gel filtration (e.g., Sephadex G-25) or dialysis against buffer. Considerations 1. Solubility: Standard Cyanine7.5 amine is hydrophobic; ensure enough co-solvent (DMSO/DMF) is used. 2. pH: The amine group must be deprotonated to react, making a slightly basic pH (8.3-8.5) ideal. 3. Applications: Ideal for near-infrared imaging (788 nm excitation/808 nm emission) of live animals.
References:
[1]. Retout M, et al. Bifunctional Calix[4]arene-Coated Gold Nanoparticles for Orthogonal Conjugation. Langmuir. 2022;38(30):9301-9309.
In Vitro, Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.) 1. Preparation of Reagents: (1) Dye Solution: Dissolve Cyanine7.5 amine in dry DMF or DMSO to create a 10-50 mg/mL stock solution. (2) Biomolecule Solution: Dissolve the protein/antibody in a coupling buffer (e.g., 50 mM borate buffer or PBS, pH 8.3-8.5) at a concentration of 1-10 mg/mL. 2. Reaction Procedure: (1) Slowly add the Cyanine7.5 amine stock solution into the protein solution while stirring. Use a molar excess of dye (typically 5-20 fold relative to the protein). (2) Incubate the mixture at room temperature for 1-2 hours or at 4°C overnight, protected from light. 3. Purification: Remove unreacted dye using gel filtration (e.g., Sephadex G-25) or dialysis against buffer. Considerations 1. Solubility: Standard Cyanine7.5 amine is hydrophobic; ensure enough co-solvent (DMSO/DMF) is used. 2. pH: The amine group must be deprotonated to react, making a slightly basic pH (8.3-8.5) ideal. 3. Applications: Ideal for near-infrared imaging (788 nm excitation/808 nm emission) of live animals.
References:
[1]. Retout M, et al. Bifunctional Calix[4]arene-Coated Gold Nanoparticles for Orthogonal Conjugation. Langmuir. 2022;38(30):9301-9309.