Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Construction and characterization of the FEMMAN chip. e, Fluorescence images of Cy5 on pGOLD substrate at (1) 200 pM, (2) 20 pM, (3) 2 pM, and (4) 200 fM compared to a quartz substrate at (5) 200 pM and (6) 20 pM. The Cy5 fluorophore density at 20 pM on both the (7) plasmonic substrate and (8) the quartz substrate was comparable. Cy5 (Glpbio) labelled primer. The reagents and DNA chip were stored in room temperature for 0 day, 7 days, 14 days, 21 days, and 30 days. Nat Biomed Eng 7.12 (2023): 1636-1648. PMID: 37735541 IF: 28.1

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This plan only provides a guide, please modify it to meet your specific needs.1. Solution preparation(1) Working solution: Dissolve CY5 solid in DMSO to a final concentration of 10 mM and filter sterilize.Note: The working solution must be prepared and used immediately. Before use, 500μg/mL condensation solution (Cat. No.: GA11126) must be used for activation before subsequent labeling experiments can be performed. 2. Experimental steps for labeling proteins with CY5(1) Protein preparation: Dissolve the protein in a pH 8.5 buffer without primary amines (such as Tris or glycine) and ammonium ions, and prepare the concentration to 2-10 mg/mL to obtain the best labeling efficiency. If the pH is lower than 8.0, adjust with 1 M sodium bicarbonate.(2) Calculation of the amount of CY5 working solution: The amount of CY5 (MW=656.81) required for the labeling reaction depends on the amount of protein to be labeled. The optimal molar ratio of CY dye to protein is about 10:1.(3) Labeling reaction: Take the calculated volume of CY5 working solution and slowly add it to the protein sample solution, gently shake to mix, and then briefly centrifuge to collect the sample at the bottom of the reaction tube. Avoid violent mixing to prevent the protein sample from denaturing and inactivating.(4) Incubation reaction: Place the reaction tube in a dark place and gently shake and incubate for 60 minutes at room temperature. Every 10-15 minutes, gently invert the reaction tube several times to fully mix the two reactants and improve the labeling efficiency. Note: (1)This product is an unactivated fluorescent dye. To use this product to label biological molecules such as peptides and proteins, it must first be activated with carboxylic acid. If an activated form is required, sulfo-Cyanine5 NHS Ester (Cat. No.: GC59174) is recommended.(2)This protocol provides guidance for using CY5 for protein labeling experiments. It can be adjusted according to other literature and specific experimental requirements. The operation should be carried out in a sterile environment to prevent contaminants from interfering with the reaction. Avoid direct contact with the reaction reagents.

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References:[1] Ferri G L, Isola J, Berger P, et al. Direct eye visualization of Cy5 fluorescence for immunocytochemistry and in situ hybridization[J]. Journal of Histochemistry & Cytochemistry, 2000, 48(3): 437-444.[2] Yuan L, Lin W, Zheng K, et al. Far-red to near infrared analyte-responsive fluorescent probes based on organic fluorophore platforms for fluorescence imaging[J]. Chemical Society Reviews, 2013, 42(2): 622-661.[3] Gerowska M, Hall L, Richardson J, et al. Efficient reverse click labeling of azide oligonucleotides with multiple alkynyl Cy-Dyes applied to the synthesis of HyBeacon probes for genetic analysis[J]. Tetrahedron, 2012, 68(3): 857-864.[4] Yu H, Chao J, Patek D, et al. Cyanine dye dUTP analogs for enzymatic labeling of DNA probes[J]. Nucleic Acids Research, 1994, 22(15): 3226-3232.[5] Ntziachristos V, Schellenberger E A, Ripoll J, et al. Visualization of antitumor treatment by means of fluorescence molecular tomography with an annexin V–Cy5. 5 conjugate[J]. Proceedings of the National Academy of Sciences, 2004, 101(33): 12294-12299.

CY5 is a cyanine-containing fluorescent dye that is commonly used to label proteins, antibodies, and small molecule compounds. Its maximum excitation/emission wavelengths are 650/670nm, respectively.

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

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CY5 is a cyanine-containing fluorescent dye that is commonly used to label proteins, antibodies, and small molecule compounds. Its maximum excitation/emission wavelengths are 650/670nm, respectively[1]. The basic structure of CY5 is a cyclic compound with a conjugated double bond system. It has excellent fluorescence properties in the far-red region and good photostability[2]. CY5 usually exists in the form of its active ester or azide, which enables it to be labeled with biological molecules such as proteins and nucleic acids through covalent bonds[3]. The optimal molar ratio of CY5 to protein is about 10:1[4]. CY5 has a wide range of applications. It is not only suitable for biomarkers and cell imaging, but also for nucleic acid detection, protein analysis, and drug screening[5]. The molecular weight of this product is 656.81. It is for research purposes only and is not suitable for food, drugs, medical devices, or cosmetics.

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Note: 1. Please make sure the liquid is clear before adding the next solvent. 2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving. 3. All of the above co-solvents are available for purchase on the GlpBio website.