Alexa Fluor 647 NHS Ester (Succinimidyl Ester)

货号：A20006,A37573,A37566,A20106

规格：1 mg,3 x 100 µg,25 mg,5 mg

价格：5988,3386,65206,23864

产品类型：蛋白标记

品牌：Thermo Fisher

Alexa Fluor 647 is a bright and photostable far-red dye with excitation ideally suited to the 633 nm laser line. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor 647 dye is water soluble and pH-insensitive from pH 4 to pH 10. Fluorescence of this long-wavelength Alexa Fluor dye is not visible to the human eye but is readily detected by most imaging systems. In addition to reactive dye formulations, we offer Alexa Fluor 647 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates optimized for cellular labeling and detection.The NHS ester (or succinimidyl ester) of Alexa Fluor 647 is the most popular tool for conjugating this dye to a protein or antibody. NHS esters can be used to label to the primary amines (R-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting Alexa Fluor conjugate will exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores.Detailed information about this AlexaFluor NHS ester:Fluorophore label:Alexa Fluor 647 dyeReactive group:NHS esterReactivity:Primary amines on proteins and ligands, amine-modified oligonucleotidesEx/Em of the conjugate:651/672 nmExtinction coefficient:270,000 cm-1M-1Spectrally similar dyes:Cy5Molecular weight:~1250Typical Conjugation ReactionYou can conjugate amine-reactive reagents with virtually any protein or peptide (the provided protocol is optimized for IgG antibodies). You can scale the reaction for any amount of protein, but the concentration of the protein should be at least 2 mg/mL for optimal results. We recommend trying three different degrees of labeling, using three different molar ratios of the reactive reagent to protein.The Alexa Fluor NHS ester is typically dissolved in high-quality anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO) (D12345), and the reaction is carried out in 0.1–0.2 M sodium bicarbonate buffer, pH 8.3, at room temperature for 1 hour. Because the pKa of the terminal amine is lower than that of the lysine epsilon-amino group, you may achieve more selective labeling of the amine terminus using a buffer closer to neutral pH.Conjugate PurificationLabeled antibodies are typically separated from free Alexa Fluor® dye using a gel filtration column, such as Sephadex G-25, BioGel P-30, or equivalent. For much larger or smaller proteins, select a gel filtration media with an appropriate molecular weight cut-off or purify by dialysis. We offer several purification kits optimized for different quantities of antibody conjugate:Antibody Conjugate Purification Kit for 0.5-1 mg (A33086)Antibody Conjugate Purification Kit for 20-50 µg (A33087)Antibody Conjugate Purification kit for 50-100 µg (A33088)

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