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  3. CY5-SE triethylamine salt

CY5-SE triethylamine salt  (Synonyms: Cy5 NHS Ester triethylamine salt; Sulfo-Cyanine5 Succinimidyl Ester triethylamine salt)

Cat. No.: HY-D0819A Purity: ≥98.0%
Handling Instructions Technical Support

Cy5-SE (Cy5 NHS Ester) triethylamine salt is a reactive dye for the labeling of amino-groups in peptides, proteins, and oligonucleotides. Cy5-SE triethylamine salt is ideal for very cost-efficient labeling of soluble proteins, as well as all kinds of peptides and oligonucleotides Ex=649 nm; Em=670 nm).

For research use only. We do not sell to patients.

CY5-SE triethylamine salt Chemical Structure

CY5-SE triethylamine salt Chemical Structure

CAS No. : 1497420-70-8

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Customer Review

Based on 16 publication(s) in Google Scholar

Other Forms of CY5-SE triethylamine salt:

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  • Biological Activity

  • Purity & Documentation

  • References

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Description

Cy5-SE (Cy5 NHS Ester) triethylamine salt is a reactive dye for the labeling of amino-groups in peptides, proteins, and oligonucleotides. Cy5-SE triethylamine salt is ideal for very cost-efficient labeling of soluble proteins, as well as all kinds of peptides and oligonucleotides Ex=649 nm; Em=670 nm)[1].

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
Add anhydrous DMSO into the vial of CY5-SE triethylamine salt to make a 10 mM stock solution. Mix well by pipetting or vortex.
3.Calculation of dye dosage
The amount of CY5-SE triethylamine salt required for reaction depends on the amount of protein to be labeled, and the optimal molar ratio of CY5-SE triethylamine salt 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-SE triethylamine salt, the required CY5-SE triethylamine salt volume is 5.73 μL, and the detailed calculation process is 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-SE triethylamine salt) = mmol (IgG) × 10 = 6.7×10-6 mmol×10 = 6.7 × 10-5 mmol
3) μL (CY5-SE triethylamine salt) = mmol (CY5-SE triethylamine salt) ×MW (CY5-SE triethylamine salt)/mg/μL (CY5-SE triethylamine salt) = 6.7 ×10-5 mmol ×855.07 mg/mmol / 0.01 mg/μL = 5.73 μL (CY5-SE triethylamine salt)
4.Run conjugation reaction
1) A good volume of freshly prepared 10 mg/mL CY5-SE triethylamine salt 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.

Molecular Weight

855.07

Formula

C43H58N4O10S2

CAS No.
Appearance

Solid

Color

Purple to purplish red

Emission (Em)

670

Excitation (Ex)

645

SMILES

O=C(CCCCCN1C2=CC=C(S(=O)(O)=O)C=C2C(C)(C)/C1=C\C=C\C=C\C3=[N+](CC)C4=C(C3(C)C)C=C(S(=O)([O-])=O)C=C4)ON5C(CCC5=O)=O.CCN(CC)CC

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-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)

Solvent & Solubility
In Vitro: 

DMSO : 125 mg/mL (146.19 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : ≥ 5.88 mg/mL (6.88 mM)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.1695 mL 5.8475 mL 11.6949 mL
5 mM 0.2339 mL 1.1695 mL 2.3390 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, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* 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    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (2.43 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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:  Saline

    Solubility: 25 mg/mL (29.24 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, 6 months; -20°C, 1 month (sealed storage, away from moisture and 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: ≥98.0%

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, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 1.1695 mL 5.8475 mL 11.6949 mL 29.2374 mL
5 mM 0.2339 mL 1.1695 mL 2.3390 mL 5.8475 mL
DMSO 10 mM 0.1169 mL 0.5847 mL 1.1695 mL 2.9237 mL
15 mM 0.0780 mL 0.3898 mL 0.7797 mL 1.9492 mL
20 mM 0.0585 mL 0.2924 mL 0.5847 mL 1.4619 mL
25 mM 0.0468 mL 0.2339 mL 0.4678 mL 1.1695 mL
30 mM 0.0390 mL 0.1949 mL 0.3898 mL 0.9746 mL
40 mM 0.0292 mL 0.1462 mL 0.2924 mL 0.7309 mL
50 mM 0.0234 mL 0.1169 mL 0.2339 mL 0.5847 mL
60 mM 0.0195 mL 0.0975 mL 0.1949 mL 0.4873 mL
80 mM 0.0146 mL 0.0731 mL 0.1462 mL 0.3655 mL
100 mM 0.0117 mL 0.0585 mL 0.1169 mL 0.2924 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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  • 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.

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CY5-SE triethylamine salt
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