BODIPY 558/568 NHS Ester (Succinimidyl Ester)/BODIPY 558/568 NHS酯（琥珀酰亚胺酯）

货号：D2219

规格：5 mg

价格：6292

产品类型：蛋白标记

品牌：Thermo Fisher

BODIPY 558/568 dye is bright, red fluorescent dye. It has a high extinction coefficient and fluorescence quantum yield and is relatively insensitive to solvent polarity and pH change. In contrast to the highly water soluble fluorophores Alexa Fluor 488 dye and fluorescein (FITC), BODIPY dyes have unique hydrophobic properties ideal for staining lipids, membranes, and other lipophilic compounds. BODIPY 558/568 dye has a relatively long excited-state lifetime (typically 5 nanoseconds or longer), which is useful for fluorescence polarization-based assays and a large two-photon cross-section for multiphoton excitation. In addition to reactive dye formulations, we offer BODIPY 558/568 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 BODIPY 558/568 is the most popular tool for conjugating the dye to a protein or antibody. NHS esters can be used to label the primary amines (R-NH2) of proteins, amine-modified oligonucleotides, and other amine-containing molecules. The resulting BODIPY 558/568 conjugates exhibit bright fluorescence, narrow emission bandwidths, and relatively long excited-state lifetimes, which can be useful for fluorescence polarization assays and two-photon excitation (TPE) microscopy.This reactive dye contains a C3alkyl spacer between the fluorophore and the NHS ester group. This spacer helps to separate the fluorophore from its point of attachment, potentially reducing the interaction of the fluorophore with the biomolecule to which it is conjugated.Detailed information about this BODIPY 558/568 NHS ester:Fluorophore label:BODIPY 558/568 dyeReactive group:NHS ester (succinimidyl ester)Reactivity:Primary amines on proteins and ligands, amine-modified oligonucleotidesEx/Em of the conjugate:559/568 nmExtinction coefficient:97,000 cm-1M-1Molecular weight:443.23Typical Conjugation ReactionAmine-reactive reagents can be conjugated with virtually any protein or peptide; the provided protocol is optimized for IgG antibodies. The reaction can be scaled 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 BODIPY NHS ester is typically dissolved in high-quality anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO), 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 BODIPY 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 medium 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)Learn More About Protein and Antibody LabelingWe offer a wide selection of Molecular Probes antibody and protein labeling kits to fit your starting material and your experimental setup. See ourAntibody Labeling kitsor use ourLabeling Chemistry Selection Toolfor other choices. To learn more about our labeling kits, readKits for Labeling Proteins and Nucleic Acids—Section 1.2in The Molecular Probes Handbook.We’ll Make a Custom Conjugate for YouIf you can’t find what you’re looking for in our online catalog, we’ll prepare a custom antibody or protein conjugate for you. Ourcustom conjugation serviceis efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

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