6-FAM  (Synonyms: 6-Carboxyfluorescein)

6-FAM (6-Carboxyfluorescein) is an isomer of carboxyfluorescein and is mainly used for sequencing and labeling of nucleic acids.

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
In cell experiments, oligonucleotide probes labeled with 6-carboxyfluorescein (6-FAM) were used, mainly through the fluorescence in situ hybridization (FISH) technique [1]. (1) Sample preparation: Prepare chromosome samples of chicken embryonic cells, including steps such as colchicine treatment, hypotonic swelling, and methanol-acetic acid fixation.
(2) Pre-treatment: The chromosomes were treated with RNase A, pepsin and formaldehyde.
(3) Hybridization: The 6-FAM-labeled oligonucleotide was dissolved in the hybridization buffer containing Escherichia coli tRNA (50% formamide, 2×SSC, 10% dextran sulfate) at a concentration of 20 ng/μL. After the DNA and the probe were denatured together, they were hybridized at room temperature in a humid box overnight.
(4) Washing and re-staining: The hybridized slides were washed three times successively in 2×SSC, then dehydrated using a series of 70%-96% ethanol, dried, and sealed with an anti-quenching agent containing DAP I.
(5) Observation and Analysis: Use the Leica DM4000B fluorescence microscope equipped with a monochrome CCD camera DFC350FX and appropriate filters to observe the samples. Then, use the QFISH software to capture and process the colored images of the chromosomes.

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

376.32

C₂₁H₁₂O₇

3301-79-9

Solid

Yellow to orange

523

489

O=C1OC2(C3=C(OC4=C2C=CC(O)=C4)C=C(O)C=C3)C5=C1C=CC(C(O)=O)=C5

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light, stored under nitrogen

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

* 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, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Alexander Stepakov, et al. Modified Synthesis of 6-carboxyfluorescein (6-FAM): Application to Probe Labeling for Conventional Cytogenetics. British Journal of Applied Science & Technology. 7(4): 423-428, 2015, Article no.BJAST.2015.160.