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  3. 5(6)-Carboxyfluorescein

5(6)-Carboxyfluorescein  (Synonyms: 5(6)-FAM; 5-(and-6)-Carboxyfluorescein mixed isomers)

Cat. No.: HY-15940 Purity: 99.51%
COA Handling Instructions

5(6)-Carboxyfluorescein (5(6)-FAM) is an amine-reactive pH-sensitive green fluorescent probe. 5(6)-Carboxyfluorescein (5(6)-FAM) can be used to label proteins, peptides and nucleotides. 5(6)-Carboxyfluorescein can be used for the detection of tumour areas in vivo.

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

5(6)-Carboxyfluorescein Chemical Structure

5(6)-Carboxyfluorescein Chemical Structure

CAS No. : 72088-94-9

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Based on 2 publication(s) in Google Scholar

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Description

5(6)-Carboxyfluorescein (5(6)-FAM) is an amine-reactive pH-sensitive green fluorescent probe. 5(6)-Carboxyfluorescein (5(6)-FAM) can be used to label proteins, peptides and nucleotides. 5(6)-Carboxyfluorescein can be used for the detection of tumour areas in vivo[1][2].

In Vitro

5(6)-Carboxyfluorescein has two main characteristics: it has two wavelengths of maximum absorbance (465 and 490 nm) and its fluorescence emission (maximum, 515 nm) increases as a function of pH in the physiological pH range of 6-7.4[2].

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

In Vivo

5(6)-Carboxyfluorescein (5 mg/kg; i.p.) can be used for in vivo pH mapping of tumor tissue[2].

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

Animal Model: CDF mice, bearing lymphoid leukemia P388[2]
Dosage: 5 mg/kg
Administration: Injected intraperitoneally
Result: Could be used for measurement and imaging of tumor tissue.
Molecular Weight

752.63

Formula

C42H24O14

CAS No.
Appearance

Solid

Color

Yellow to orange

Emission (Em)

515

Excitation (Ex)

488

SMILES

OC1=CC=C(C2(O3)C(C=C(C(O)=O)C=C4)=C4C3=O)C(OC5=C2C=CC(O)=C5)=C1.OC6=CC=C(C7(O8)C(C=CC(C(O)=O)=C9)=C9C8=O)C(OC%10=C7C=CC(O)=C%10)=C6

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 : 62.5 mg/mL (83.04 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 1.3287 mL 6.6434 mL 13.2867 mL
5 mM 0.2657 mL 1.3287 mL 2.6573 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
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Concentration
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Volume
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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 (2.76 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 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.
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: 99.51%

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 1.3287 mL 6.6434 mL 13.2867 mL 33.2169 mL
5 mM 0.2657 mL 1.3287 mL 2.6573 mL 6.6434 mL
10 mM 0.1329 mL 0.6643 mL 1.3287 mL 3.3217 mL
15 mM 0.0886 mL 0.4429 mL 0.8858 mL 2.2145 mL
20 mM 0.0664 mL 0.3322 mL 0.6643 mL 1.6608 mL
25 mM 0.0531 mL 0.2657 mL 0.5315 mL 1.3287 mL
30 mM 0.0443 mL 0.2214 mL 0.4429 mL 1.1072 mL
40 mM 0.0332 mL 0.1661 mL 0.3322 mL 0.8304 mL
50 mM 0.0266 mL 0.1329 mL 0.2657 mL 0.6643 mL
60 mM 0.0221 mL 0.1107 mL 0.2214 mL 0.5536 mL
80 mM 0.0166 mL 0.0830 mL 0.1661 mL 0.4152 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)-Carboxyfluorescein
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