Determine the necessary mass, volume, or concentration for preparing a solution.
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™ 647 is a bright, far-red fluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. 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. 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 (learn more). The maleimide derivative of Alexa Fluor™ 647 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™ 647 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™ 647 dye Reactive group: maleimide Reactivity: thiol groups on proteins and ligands, oligonucleotide thiophosphates Ex/Em of the conjugate: 651/671 nm Extinction coefficient: 265,000 cm-1M-1 Spectrally similar dyes: Cy5 Molecular weight: ∼1250 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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Enter Lot Number to search for COA:
1. Molina ML, Barrera FN, Fernández AM, Poveda JA, Renart ML, Encinar JA, Riquelme G, González-Ros JM. () Clustering and coupled gating modulate the activity in KcsA, a potassium channel model.. J Biol Chem, [PMID:16670090] |
2. Wesolowska N, Avilov I, Machado P, Geiss C, Kondo H, Mori M, Lenart P. () Actin assembly ruptures the nuclear envelope by prying the lamina away from nuclear pores and nuclear membranes in starfish oocytes.. Elife, [PMID:31989921] |
3. Wheelock AM, Morin D, Bartosiewicz M, Buckpitt AR. () Use of a fluorescent internal protein standard to achieve quantitative two-dimensional gel electrophoresis.. Proteomics, [PMID:16429456] |
4. Palchevskyy SS, Posokhov YO, Olivier B, Popot JL, Pucci B, Ladokhin AS. () Chaperoning of insertion of membrane proteins into lipid bilayers by hemifluorinated surfactants: application to diphtheria toxin.. Biochemistry, [PMID:16489756] |
5. Kuzmenkina EV, Heyes CD, Nienhaus GU. () Single-molecule Forster resonance energy transfer study of protein dynamics under denaturing conditions.. Proc Natl Acad Sci U S A, [PMID:16221762] |
6. Jäger M, Michalet X, Weiss S. () Protein-protein interactions as a tool for site-specific labeling of proteins.. Protein Sci, [PMID:15987886] |
7. Ladokhin AS, Haigler HT. () Reversible transition between the surface trimer and membrane-inserted monomer of annexin 12.. Biochemistry, [PMID:15736950] |
8. Bowen ME, Weninger K, Ernst J, Chu S, Brunger AT. () Single-molecule studies of synaptotagmin and complexin binding to the SNARE complex.. Biophys J, [PMID:15821166] |
9. Patel DR, Isas JM, Ladokhin AS, Jao CC, Kim YE, Kirsch T, Langen R, Haigler HT. () The conserved core domains of annexins A1, A2, A5, and B12 can be divided into two groups with different Ca2+-dependent membrane-binding properties.. Biochemistry, [PMID:15723527] |
10. Weninger K, Bowen ME, Chu S, Brunger AT. () Single-molecule studies of SNARE complex assembly reveal parallel and antiparallel configurations.. Proc Natl Acad Sci U S A, [PMID:14657376] |
11. Yang W, Musser SM. () Nuclear import time and transport efficiency depend on importin beta concentration.. J Cell Biol, [PMID:16982803] |
12. Kong X, Nir E, Hamadani K, Weiss S,. () Photobleaching pathways in single-molecule FRET experiments.. J Am Chem Soc, [PMID:17375921] |
13. Ramachandiran V, Grigoriev V, Lan L, Ravkov E, Mertens SA, Altman JD. () A robust method for production of MHC tetramers with small molecule fluorophores.. J Immunol Methods, [PMID:17187819] |
14. Stanoev A, Mhamane A, Schuermann KC, Grecco HE, Stallaert W, Baumdick M, Brüggemann Y, Joshi MS, Roda-Navarro P, Fengler S, Stockert R, Roßmannek L, Luig J, Koseska A, Bastiaens PIH. () Interdependence between EGFR and Phosphatases Spatially Established by Vesicular Dynamics Generates a Growth Factor Sensing and Responding Network.. Cell Syst, [PMID:30145116] |
15. Trelle MB, Ramsey KM, Lee TC, Zheng W, Lamboy J, Wolynes PG, Deniz A, Komives EA. () Binding of NF?B Appears to Twist the Ankyrin Repeat Domain of I?Ba.. Biophys J, [PMID:26910425] |
16. Chandrasekaran R, Kenworthy AK, Lacy DB. () Clostridium difficile Toxin A Undergoes Clathrin-Independent, PACSIN2-Dependent Endocytosis.. PLoS Pathog, [PMID:27942025] |
17. Pesenti ME, Prumbaum D, Auckland P, Smith CM, Faesen AC, Petrovic A, Erent M, Maffini S, Pentakota S, Weir JR, Lin YC, Raunser S, McAinsh AD, Musacchio A. () Reconstitution of a 26-Subunit Human Kinetochore Reveals Cooperative Microtubule Binding by CENP-OPQUR and NDC80.. Mol Cell, [PMID:30174292] |
18. Chung HS, Meng F, Kim JY, McHale K, Gopich IV, Louis JM. () Oligomerization of the tetramerization domain of p53 probed by two- and three-color single-molecule FRET.. Proc Natl Acad Sci U S A, [PMID:28760960] |
19. Cedeño C, Pauwels K, Tompa P. () Protein Delivery into Plant Cells: Toward. Front Plant Sci, [PMID:28469623] |
20. Zamir E, Frey C, Weiss M, Antona S, Frohnmayer JP, Janiesch JW, Platzman I, Spatz JP. () Reconceptualizing Fluorescence Correlation Spectroscopy for Monitoring and Analyzing Periodically Passing Objects.. Anal Chem, [PMID:28985462] |
21. DeRocco V, Anderson T, Piehler J, Erie DA, Weninger K,. () Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes.. Biotechniques, [PMID:21091445] |
22. Alam IS, Neves AA, Witney TH, Boren J, Brindle KM,. () Comparison of the C2A domain of synaptotagmin-I and annexin-V as probes for detecting cell death.. Bioconjug Chem, [PMID:20402461] |
23. Gambin Y, Deniz AA,. () Multicolor single-molecule FRET to explore protein folding and binding.. Mol Biosyst, [PMID:20601974] |
24. McCann JJ, Choi UB, Zheng L, Weninger K, Bowen ME,. () Optimizing methods to recover absolute FRET efficiency from immobilized single molecules.. Biophys J, [PMID:20682275] |
25. Zhang Y, Sivasankar S, Nelson WJ, Chu S,. () Resolving cadherin interactions and binding cooperativity at the single-molecule level.. Proc Natl Acad Sci U S A, [PMID:19114658] |
26. Lu Y,Wang W,Kirschner MW. () Specificity of the anaphase-promoting complex: a single-molecule study.. , [PMID:25859049] |
27. Das SK, Darshi M, Cheley S, Wallace MI, Bayley H. () Membrane protein stoichiometry determined from the step-wise photobleaching of dye-labelled subunits.. Chembiochem, [PMID:17503420] |
28. Tessier PM, Lindquist S. () Prion recognition elements govern nucleation, strain specificity and species barriers.. Nature, [PMID:17495929] |
29. Majumdar DS, Smirnova I, Kasho V, Nir E, Kong X, Weiss S, Kaback HR,. () Single-molecule FRET reveals sugar-induced conformational dynamics in LacY.. Proc Natl Acad Sci U S A, [PMID:17502603] |
30. Wendorff TJ, Berger JM. () Topoisomerase VI senses and exploits both DNA crossings and bends to facilitate strand passage.. Elife, [PMID:29595473] |
31. Maxwell D, Chang Q, Zhang X, Barnett EM, Piwnica-Worms D,. () An improved cell-penetrating, caspase-activatable, near-infrared fluorescent peptide for apoptosis imaging.. Bioconjug Chem, [PMID:19331388] |
32. Peng W, Shi J, Márquez CL, Lau D, Walsh J, Faysal KMR, Byeon CH, Byeon IL, Aiken C, Böcking T. () Functional analysis of the secondary HIV-1 capsid binding site in the host protein cyclophilin A.. Retrovirology, [PMID:30947724] |
33. Vahey MD, Fletcher DA. () Influenza A virus surface proteins are organized to help penetrate host mucus.. Elife, [PMID:31084711] |
34. Duan Y, Du A, Gu J, Duan G, Wang C, Gui X, Ma Z, Qian B, Deng X, Zhang K, Sun L, Tian K, Zhang Y, Jiang H, Liu C, Fang Y. () PARylation regulates stress granule dynamics, phase separation, and neurotoxicity of disease-related RNA-binding proteins.. Cell Res, [PMID:30728452] |
35. Sparks JL, Chistol G, Gao AO, Räschle M, Larsen NB, Mann M, Duxin JP, Walter JC. () The CMG Helicase Bypasses DNA-Protein Cross-Links to Facilitate Their Repair.. Cell, [PMID:30595447] |
36. Chen Y, Yong J, Martínez-Sánchez A, Yang Y, Wu Y, De Camilli P, Fernández-Busnadiego R, Wu M. () Dynamic instability of clathrin assembly provides proofreading control for endocytosis.. J Cell Biol, [PMID:31451612] |
37. Scarborough EA, Davis TN, Asbury CL. () Tight bending of the Ndc80 complex provides intrinsic regulation of its binding to microtubules.. Elife, [PMID:31045495] |
38. Kim Y, Ho SO, Gassman NR, Korlann Y, Landorf EV, Collart FR, Weiss S,. () Efficient site-specific labeling of proteins via cysteines.. Bioconjug Chem, [PMID:18275130] |
39. Orte A, Craggs TD, White SS, Jackson SE, Klenerman D,. () Evidence of an intermediate and parallel pathways in protein unfolding from single-molecule fluorescence.. J Am Chem Soc, [PMID:18507381] |
40. Andrecka J, Lewis R, Brückner F, Lehmann E, Cramer P, Michaelis J,. () Single-molecule tracking of mRNA exiting from RNA polymerase II.. Proc Natl Acad Sci U S A, [PMID:18162559] |