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Storage Temp | Protected from light,Store at -20°C |
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Shipped In | Ice chest + Ice pads |
Product Description | Product introduction: Alexa Fluor™ 568 is a bright, orange/red fluorescent dye with excitation ideally suited for the 568 nm laser line on the Ar-Kr mixed-gas laser. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor™ 568 dye is water soluble and pH-insensitive from pH 4 to pH 10. In addition to reactive dye formulations, we offer Alexa Fluor™ 568 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates optimized for cellular labeling and detection (learn more). The maleimide derivative of Alexa Fluor™ 568 is the most popular tool for conjugating the dye to a thiol group on a protein, oligonucleotide thiophosphate, or low molecular weight ligand. The resulting Alexa Fluor™ 568 conjugates exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores. Detailed information about this AlexaFluor™ maleimide: Fluorophore label: Alexa Fluor™ 568 dye Reactive group: maleimide Reactivity: thiol groups on proteins and ligands, oligonucleotide thiophosphates Ex/Em of the conjugate: 575/600 nm Extinction coefficient: 92,000 cm-1M-1 Spectrally similar dyes: Rhodamine red Molecular weight: 880.92 Typical Conjugation Reaction The protein should be dissolved at a concentration of 50-100 µM in a suitable buffer (10-100 mM phosphate, Tris, or HEPES) at pH 7.0-7.5. In this pH range, the protein thiol groups are sufficiently nucleophilic that they react almost exclusively with the reagent in the presence of the more numerous protein amine groups, which are protonated and relatively unreactive. We recommend reducing any disulfide bonds at this point using a 10-fold molar excess of reducing agent such as DTT or TCEP. Excess DTT must be removed by dialysis and subsequent thiol-modification should be carried out under oxygen-free conditions to prevent reformation of the disulfide bonds; these precautions are not necessary when using TCEP prior to maleimide conjugation. The Alexa Fluor™ maleimide is typically dissolved in high-quality anhydrous dimethylsulfoxide (DMSO) at a concentration of 1-10 mM immediately prior to use, and stock solutions should be protected from light as much as possible. Generally, this stock solution is added to the protein solution dropwise while stirring to produce approximately 10-20 moles of reagent per mole of protein, and the reaction is allowed to proceed at room temperature for 2 hours or at 4°C overnight, protected from light. Any unreacted thiol-reactive reagent can be consumed by adding excess glutathione, mercaptoethanol, or other soluble low molecular weight thiol. Conjugate Purification Labeled 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 () Antibody Conjugate Purification Kit for 20-50 µg () Antibody Conjugate Purification kit for 50-100 µg () Learn More About Protein and Antibody Labeling We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See ourAntibody Labeling kits or use ourKits for Labeling Proteins and Nucleic Acids—Section 1.2 in The Molecular Probes™ Handbook. We’ll Make a Custom Conjugate for You If 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 service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified. |
Reactivity | Thiol |
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1. Kästner CN, Prummer M, Sick B, Renn A, Wild UP, Dimroth P. () The citrate carrier CitS probed by single-molecule fluorescence spectroscopy.. Biophys J, [PMID:12609868] |
2. de Diesbach P,N'Kuli F,Berens C,Sonveaux E,Monsigny M,Roche AC,Courtoy PJ. () Receptor-mediated endocytosis of phosphodiester oligonucleotides in the HepG2 cell line: evidence for non-conventional intracellular trafficking.. Nucleic Acids Res, [PMID:11917011] |
3. Marras SA, Kramer FR, Tyagi S. () Efficiencies of fluorescence resonance energy transfer and contact-mediated quenching in oligonucleotide probes.. Nucleic Acids Res, [PMID:12409481] |
4. Martin AC, Welch MD, Drubin DG. () Arp2/3 ATP hydrolysis-catalysed branch dissociation is critical for endocytic force generation.. Nat Cell Biol, [PMID:16862144] |
5. Granier S, Kim S, Shafer AM, Ratnala VR, Fung JJ, Zare RN, Kobilka B,. () Structure and conformational changes in the C-terminal domain of the beta2-adrenoceptor: insights from fluorescence resonance energy transfer studies.. J Biol Chem, [PMID:17347144] |