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  4. Cy5.5

Cy5.5  (Synonyms: Sulfo-Cyanine5.5)

Cat. No.: HY-D0924 Purity: 98.19%
COA Handling Instructions

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Cy5.5 is a CY dye. CY, short for Cyanine, is a compound consisting of two nitrogen atoms connected by an odd number of methyl units. Cyanine compounds have the characteristics of long wavelength, adjustable absorption and emission, high extinction coefficient, good water solubility and relatively simple synthesis. CY dyes are of en used for the labeling of proteins, antibodies and small molecular compounds. For the labeling of protein antibodies, the combination can be completed through a simple mixing reaction. Below, we introduce the labeling method of protein antibody labeling, which has certain reference significance.

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Cy5.5 Chemical Structure

Cy5.5 Chemical Structure

CAS No. : 210892-23-2

Size Price Stock Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO
ready for reconstitution
 USD 343 In-stock
Solution
10 mM * 1 mL in DMSO USD 343 In-stock
Solid
1 mg USD 60 In-stock
5 mg USD 200 In-stock
10 mg USD 340 In-stock
25 mg USD 650 In-stock
50 mg USD 860 In-stock
100 mg USD 1120 In-stock
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Based on 23 publication(s) in Google Scholar

Other Forms of Cy5.5:

Cy5.5 Related Antibodies

Top Publications Citing Use of Products

    Cy5.5 purchased from MedChemExpress. Usage Cited in: Adv Funct Mater. 2022: 2204636.

    Representative confocal images of DC2.4 and RAW264.7 cells incubates with Cy5.5 labelled mOVA and mRLNPs for 6 h, showing cellular uptake and endo/lysosome escaping of mOVA. The endo/lysosomes are stained with LysoTracker Green (green).

    • Biological Activity

    • Purity & Documentation

    • References

    • Customer Review

    Description

    Cy5.5 is a CY dye. CY, short for Cyanine, is a compound consisting of two nitrogen atoms connected by an odd number of methyl units. Cyanine compounds have the characteristics of long wavelength, adjustable absorption and emission, high extinction coefficient, good water solubility and relatively simple synthesis[1]. CY dyes are of en used for the labeling of proteins, antibodies and small molecular compounds. For the labeling of protein antibodies, the combination can be completed through a simple mixing reaction. Below, we introduce the labeling method of protein antibody labeling, which has certain reference significance[2].

    In Vitro

    Protocol
    1.Protein Preparetion
    1) In order to obtain the best labeling effect, please prepare the protein (antibody) concentration as 2 mg/mL.
    2) The pH value of protein solution shall be 8.5±0.5. If the pH is lower than 8.0, 1 M sodium bicarbonate shall be used for adjustment.
    3) If the protein concentration is lower than 2 mg/mL, the labeling efficiency will be greatly reduced. In order to obtain the best labeling efficiency, it is recommended that the final protein concentration range is 2-10 mg/mL.
    4) The protein must be in the buffer without primary amine (such as Tris or glycine) and ammonium ion, otherwise the labeling efficiency will be affected.
    2.Dye Preparation (Cy5.5)
    Add anhydrous DMSO into the vial of Cy5.5 to make a 10 mM stock solution. Mix well by pipetting or vortex.
    Before use, it must be activated with condensation solution (500 μg/mL) (HY-D0178) before subsequent labeling experiments can be performed.
    3.Calculation of dye dosage
    The amount of Cy5.5 required for reaction depends on the amount of protein to be labeled, and the optimal molar ratio of Cy5.5 to protein is about 10.
    Example: assuming the required marker protein is 500 μL 2 mg/mL IgG (MW=150,000), use 100 μL DMSO dissolve 1 mg Cy5.5, the required Cy5.5 volume calculation process as follows:
    1) mmol (IgG) = mg/mL (IgG) ×mL (IgG) / MW (IgG) =2 mg/mL × 0.5 mL / 150,000 mg/mmol= 6.7×10-6 mmol
    2) mmol (Cy5.5) = mmol (IgG) × 10 = 6.7×10-6 mmol×10 = 6.7 × 10-5 mmol
    3) μL (Cy5.5) = mmol (Cy5.5) ×MW (Cy5.5) / mg/μL (Cy5.5) = 6.7 ×10-5 mmol ×917.05 mg/mmol / 0.01 mg/μL
    4.Run conjugation reaction
    1) A calculated volume of freshly prepared 10 mM Cy5.5 (50 μL of 500 μg/mL condensation solution can be used to activate about 10 μL dye solution) is slowly added to 0.5 mL protein sample
    In solution, gently shake to mix, then centrifuge briefly to collect the sample at the bottom of the reaction tube. Don'tmix well to prevent protein samples from denaturation and inactivation.
    2) The reaction tubules were placed in a dark place and incubated gently at room temperature for 60 minutes at intervals.For 10-15 minutes, gently reverse the reaction tubules several times to fully mix the two reactants and raise the bar efficiency.
    5.Purify the conjugation
    The following protocol is an example of dye-protein conjugate purification by using a SepHadex G-25 column.
    1) Prepare SepHadex G-25 column according to the manufacture instruction.
    2) Load the reaction mixture (From "Run conjugation reaction") to the top of the SepHadex G-25 column.
    3) Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface.
    4) Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate.

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

    Cy5.5 Related Antibodies
    In Vivo

    Cy5.5-labeled factor VIIa is developed for imagining cancer. Cy5.5 labeled with these targeting proteins specifically localize to the tumor xenografts for at least 14 days but unconjugated Cy5.5 does not localize to any xenografts or organs. This method of imaging anti-tissue factor in the tumor VECs may be useful in detecting primary tumors and metastases as well as monitoring in vivo therapeutic responses[1]. pH/temperature sensitive magnetic nanogels conjugated with Cy5.5-labled lactoferrin (Cy5.5-Lf-MPNA nanogels) are developed as a promising contrast agent for preoperative MRI and intraoperative fluorescence imaging of glioma[2].

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
    Molecular Weight

    917.05

    Formula

    C41H44N2O14S4
    CAS No.
    Appearance

    Solid

    Color

    Purple to purplish red

    Emission (Em)

    710
    Excitation (Ex)

    680
    SMILES

    O=S(C1=CC(S(=O)([O-])=O)=C2C(C3=C([N+](CC)=C(/C=C/C=C/C=C4N(CCCCCC(O)=O)C5=C(C6=CC(S(=O)(O)=O)=CC(S(=O)(O)=O)=C6C=C5)C/4(C)C)C3(C)C)C=C2)=C1)(O)=O
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    4°C, protect from light

    *In solvent : -80°C, 2 years; -20°C, 1 year (protect from light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 100 mg/mL (109.05 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    H2O : 5 mg/mL (5.45 mM; ultrasonic and warming and heat to 60°C)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.0905 mL 5.4523 mL 10.9045 mL
    5 mM 0.2181 mL 1.0905 mL 2.1809 mL
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year (protect from light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

    • Molarity Calculator

    • Dilution Calculator

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
    =
    Concentration
    ×
    Volume
    ×
    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 4.17 mg/mL (4.55 mM); Clear solution

      This protocol yields a clear solution of ≥ 4.17 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (41.7 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
    • Protocol 2

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

      Solubility: ≥ 4.17 mg/mL (4.55 mM); Clear solution

      This protocol yields a clear solution of ≥ 4.17 mg/mL (saturation unknown).

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (41.7 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

      Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

    For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  PBS

      Solubility: 5 mg/mL (5.45 mM); Clear solution; Need ultrasonic and warming and heat to 60°C

    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    % 
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

    *In solvent : -80°C, 2 years; -20°C, 1 year (protect from light)

    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation

    Purity: 99.88%

    COA (255 KB) HNMR (133 KB) LCMS (93 KB)

    Handling Instructions (2659 KB)
    Dyeing Example
    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year (protect from light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    H2O / DMSO 1 mM 1.0905 mL 5.4523 mL 10.9045 mL 27.2613 mL
    5 mM 0.2181 mL 1.0905 mL 2.1809 mL 5.4523 mL
    DMSO 10 mM 0.1090 mL 0.5452 mL 1.0905 mL 2.7261 mL
    15 mM 0.0727 mL 0.3635 mL 0.7270 mL 1.8174 mL
    20 mM 0.0545 mL 0.2726 mL 0.5452 mL 1.3631 mL
    25 mM 0.0436 mL 0.2181 mL 0.4362 mL 1.0905 mL
    30 mM 0.0363 mL 0.1817 mL 0.3635 mL 0.9087 mL
    40 mM 0.0273 mL 0.1363 mL 0.2726 mL 0.6815 mL
    50 mM 0.0218 mL 0.1090 mL 0.2181 mL 0.5452 mL
    60 mM 0.0182 mL 0.0909 mL 0.1817 mL 0.4544 mL
    80 mM 0.0136 mL 0.0682 mL 0.1363 mL 0.3408 mL
    100 mM 0.0109 mL 0.0545 mL 0.1090 mL 0.2726 mL

    * Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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    Cy5.5 Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    Cy5.5210892-23-2Sulfo-Cyanine5.5Fluorescent DyeInhibitorinhibitorinhibit

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