In the field of biochemical research and molecular labeling, the 5 FAM peptide stands out as a critical tool for scientists. As someone who has frequently utilized various fluorescent reagents for laboratory analysis, I have found that understanding the specific properties of 5-FAM (5-Carboxyfluorescein) is essential for achieving consistent, high-quality results. This article explores the tec 5-FAM lysine hnical characteristics and practical utility of this versatile molecule.
Technical Profile of 5-FAM
The 5-FAM (5-Carboxyfluorescein) molecule is a highly purified single isomer that serves as a c 5-FAM [5-Carboxyfluorescein] ornerstone in fluorescent labeling. With a molecular weight of MW 376.3 Da and a Purity >99%, it provides the precision required for delicate experiments. When observing its optical properties, researchers note that there are characteristic absorption bands of 5-FAM that make it an ideal candidate for detection, typically exhibiting an excitation maximum around 492–494 nm and an emission around 517–518 nm.
Because 5-FAM is a green fluorescent reagent used for in situ labeling peptides, proteins, and nucleotides, it has become a standard in the industry. It is a green fluorescent probe in the form of a single isomer, which ensures that the labeling process is clean and predictable, avoiding the complexities that can arise from mixed isomers like 6-FAM.
Key Applications and Functionality
My experience with these reagents highlights their adaptability. F 5-FAM is a single isomer. It is one of the most populargreen fluorescent reagents used for in situ labeling peptides, proteins and nucleotides.Read more or instance, the collagen hybridizing peptide (CHP), often con by AF Dyes—Carboxyfluorescein (FAM): It is a popular green fluorophore (excitation/emission: ~494/518 nm) used in various biological assays. · Fluorescein isothiocyanate ( Read more jugated with this fluorophore, can specifically bind to denatured collagen strands through hydrogen bonding. This is a remarkable feature for researchers studying tissue structure.
Furthermore, 5-FAM acts as an excellent building block to prepare fluorescent peptides for various assays. In my own MW 376.3 Da, Purity >99%. Amine-reactive fluorescent label. 6-FAM (ab145294) isomer. Polar tracer. Covalently labels peptides and proteins. 5-CFDA Read more observations, when working with a 5-FAM conjugate, it is important to note that you should have the protein in a non-amine buffer (such as PBS, pH 7.2–7.5) to ensure optimal conjugation.
Some specific variations include:
* 5-FAM-SE: This is the amine-reactive succinimidyl ester of single isomer 5-FAM acid, which is highly effective for stable covalent bonding.
* FAM reagents: These give carboxamides that are more resistant to hydrolysis, 5-FAM, SE (λex/λem: 492/518nm in DMF)is the amine-reactive succinimidyl ester of single isomer 5-FAM acid. It is one of the most popular green fluorescent Read more making them durable during long-term storage or experimental procedures.
* Specialized peptides: For example, Crosstide is a peptide analog of glycogen synthase kinase-3 that is often studied using these labeling techniques. Similarly, the RGD peptide is a ligand for cell-surface integrin receptors, providing a pathway for targeted studies in cellular environments.
Practical Considerations for Researchers
When preparing solutions, such as a 5 μM H3-fluorescein (H3-FAM) peptide in water, the purity of the 5-Carboxyfluorescein is paramount Absorption spectrum of 5-FAM/Cy5 peptide.Characteristic absorption bands of 5-FAMand Cy5 are clearly separated. The concentration of the peptide was 10μM.Read more . Because FAM is a carboxyfluorescein that is sensitive to pH changes, it is typically used in a range of 7.5 to 8.5 to maintain peak fluorescence.
Beyond its standard use, 5-Carboxyfluorescein also functions as a key building block for developing Big Dye DNA sequencing terminators. Whether you are working with a peptide or a complex protein, the 5 (referring to the 5-isomer position) ensures that the Fam (fluorescein) group remains stable and provides the necessary signal intensity for high-resolution imaging.
Summary
The use of 5 FAM peptide conjugates remains a gold standard in laboratories worldwide. By selecting the correct isomer and adhering to proper buffer conditions, researchers can leverage the high purity and specific binding capabilities of these molecules to advance their work in molecular biology. Whether investigating collagen hybridizing peptide (CHP) applications or performing standard protein labeling, the reliability of 5-FAM as the free acid form of 5-carboxyfluorescein single isomer continues to be an invaluable asset.