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  3. 5(6)-TAMRA

5(6)-TAMRA  (Synonyms: 5(6)-Carboxytetramethylrhodamine)

Cat. No.: HY-15944 Purity: 99.72%
Handling Instructions Technical Support

5(6)-TAMRA (5(6)-Carboxytetramethylrhodamine) is a fluorescent dye molecule widely used as a label for peptides and proteins (Ex/Em = 520/600 nm).

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

5(6)-TAMRA Chemical Structure

5(6)-TAMRA Chemical Structure

CAS No. : 98181-63-6

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Based on 2 publication(s) in Google Scholar

Other Forms of 5(6)-TAMRA:

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2 Publications Citing Use of MCE 5(6)-TAMRA

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  • Purity & Documentation

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Description

5(6)-TAMRA (5(6)-Carboxytetramethylrhodamine) is a fluorescent dye molecule widely used as a label for peptides and proteins (Ex/Em = 520/600 nm).

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 5(6)-TAMRA 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 5(6)-TAMRA required for reaction depends on the amount of protein to be labeled, and the optimal molar ratio of 5(6)-TAMRA 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 5(6)-TAMRA, the required 5(6)-TAMRA volume is 2.88 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 (5(6)-TAMRA) = mmol (IgG) × 10 = 6.7×10-6 mmol×10 = 6.7 × 10-5 mmol
3) μL (5(6)-TAMRA) = mmol (5(6)-TAMRA) ×MW (5(6)-TAMRA)/mg/μL (5(6)-TAMRA) = 6.7 ×10-5 mmol × 430.45 mg/mmol/0.01 mg/μL = 2.88 μL (5(6)-TAMRA)
4.Run conjugation reaction
1) A good volume of freshly prepared 10 mg/mL 5(6)-TAMRA 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

430.45

Formula

C25H22N2O5

CAS No.
Appearance

Solid

Color

Brown to black

Emission (Em)

600

Excitation (Ex)

520

SMILES

O=C(C1=C2C=CC(C(O)=O)=C1)OC32C(C=CC(N(C)C)=C4)=C4OC5=C3C=CC(N(C)C)=C5.O=C(C6=C7C=C(C(O)=O)C=C6)OC87C(C=CC(N(C)C)=C9)=C9OC%10=C8C=CC(N(C)C)=C%10

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

DMSO : ≥ 20.83 mg/mL (48.39 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

*"≥" means soluble, but saturation unknown.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.3232 mL 11.6158 mL 23.2315 mL
5 mM 0.4646 mL 2.3232 mL 4.6463 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • 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: 2.08 mg/mL (4.83 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 2.08 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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.
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 (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: 99.72%

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, 6 months; -20°C, 1 month (protect from 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
DMSO 1 mM 2.3232 mL 11.6158 mL 23.2315 mL 58.0788 mL
5 mM 0.4646 mL 2.3232 mL 4.6463 mL 11.6158 mL
10 mM 0.2323 mL 1.1616 mL 2.3232 mL 5.8079 mL
15 mM 0.1549 mL 0.7744 mL 1.5488 mL 3.8719 mL
20 mM 0.1162 mL 0.5808 mL 1.1616 mL 2.9039 mL
25 mM 0.0929 mL 0.4646 mL 0.9293 mL 2.3232 mL
30 mM 0.0774 mL 0.3872 mL 0.7744 mL 1.9360 mL
40 mM 0.0581 mL 0.2904 mL 0.5808 mL 1.4520 mL
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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.

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Product Name:
5(6)-TAMRA
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