Alexa fluor 647 maleimide dye

Manufactured by Thermo Fisher Scientific

Alexa Fluor 647 maleimide dyes are fluorescent labeling reagents designed for the covalent attachment of dyes to thiol-containing biomolecules. The dyes feature high molar absorption coefficients, bright fluorescence, and photostability. They are commonly used in applications such as flow cytometry, microscopy, and immunoassays.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (6 mentions) Alexa fluor 488 maleimide dye, by Thermo Fisher Scientific (2 mentions) Pd spintrap g 25 column, by Cytiva (2 mentions) Glucose oxidase, by Serva Electrophoresis (2 mentions) Catalase, by Merck Group (2 mentions) Alexa fluor 555, by Thermo Fisher Scientific (2 mentions) H2so4, by Merck Group (2 mentions) Adenosine triphosphate (atp), by Merck Group (2 mentions) Tris buffered saline (tbs), by Corning (1 mentions) Nicl2, by Merck Group (1 mentions) Tris 2 carboxyethyl phosphine tcep, by Thermo Fisher Scientific (1 mentions) Glucose and h2o2, by Thermo Fisher Scientific (1 mentions) 1 2 dioleoyl sn glycero 3 phosphocholine dopc, by Avanti Polar Lipids (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) 2 mercaptoethanol, by Merck Group (1 mentions) 6 hydroxy 2 5 7 8 tetramethylchromane 2 carboxylic acid trolox, by Merck Group (1 mentions)

Spelling variants of this product

Alexa Fluor 647 (1520 citations) Alexa 647 (424 citations) Alexa Fluor dyes (206 citations) Alexa dyes (49 citations) Alexa Fluors (21 citations) Alexa 647 maleimide (18 citations) Alexa Fluor 647 Maleimide (16 citations) Alexa 647 dye (11 citations)

5 protocols using alexa fluor 647 maleimide dye

Labeling and Disaggregation of α-Synuclein Amyloids

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S9C:WT (1:2) αSyn amyloid fibres were incubated with Alexa Fluor 647 maleimide dye (10‐fold molar excess, Thermo Fisher Scientific) and Biotin‐X, SE dye (2.5‐fold molar excess, Thermo Fisher Scientific) for at least 2 h at room temperature in the dark in HKMT buffer. T111C Hsc70 was incubated with Alexa Fluor 488 maleimide dye (10‐fold molar excess, Thermo Fisher Scientific) for at least 2 h at room temperature in the dark in HKMT buffer. Labelled Hsc70 was then buffer exchanged using PD SpinTrap G‐25 column (Cytiva) pre‐equilibrated in HKMD buffer. Labelled amyloid fibres (1 µM) were incubated with labelled Hsc70 (2 µM), DNAJB1 (1 µM) and Apg2 (0.2 µM, if indicated) in disaggregation buffer for 1 h at 30°C in the dark. The T111C‐Alexa Fluor 488 Hsc70 had the same disaggregation activity as unlabelled T111C‐Hsc70 (Fig EV5).

Beton J.G., Monistrol J., Wentink A., Johnston E.C., Roberts A.J., Bukau B.G., Hoogenboom B.W, & Saibil H.R. (2022). Cooperative amyloid fibre binding and disassembly by the Hsp70 disaggregase. The EMBO Journal, 41(16), e110410.

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Hsc70-mediated Amyloid Disaggregation

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S9C:WT (1:2) αSyn amyloid fibres were incubated with Alexa Fluor 647 maleimide dye (10-fold molar excess, Thermo Fisher Scientific) and Biotin-X, SE dye (2.5-fold molar excess, Thermo Fisher Scientific) for at least 2 hours at room temperature in the dark in HKMT buffer. T111C Hsc70 was incubated with Alexa Fluor 488 maleimide dye (10-fold molar excess, Thermo Fisher Scientific) for at least 2 hours at room temperature in the dark in HKMT buffer. Labelled Hsc70 was then buffer exchanged using PD SpinTrap G-25 column (Cytiva) pre-equilibrated in HKMD buffer. Labelled amyloid fibres (1 µM) were incubated with labelled Hsc70 (2 µM), DNAJB1 (1 µM) and Apg2 (0.2 µM, if indicated) in disaggregation buffer for 1 hour at 30°C in the dark.

Beton J.G., Monistrol J., Wentink A.S., Johnston E.F., Roberts A., Bukau B., Hoogenboom B.W., & Saibil H. (2021). Cooperative Amyloid Fibre Binding and Disassembly by the Hsp70 disaggregase.

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Labeling Proteins with Fluorescent Dyes

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DOPC and 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (Ni²⁺-NTA-DOGS; nickel salt) were purchased from Avanti Polar Lipids. Alexa Fluor 488, Alexa Fluor 555, and Alexa Fluor 647 maleimide dyes were purchased from Thermo Fisher. Bovine serum albumin (BSA), (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, catalase, 2-mercaptoethanol (BME), NiCl₂, H₂SO₄, and ATP were purchased from Sigma-Aldrich. Glucose oxidase was purchased from Serva. Tris(2-carboxyethyl)phosphine (TCEP), glucose and H₂O₂ were purchased from Thermo Fisher Scientific. MgCl₂ was purchased from EMD Chemicals. Tris-buffered saline (TBS) was purchased from Corning.

Lin C.W., Nocka L.M., Stinger B.L., DeGrandchamp J.B., Lew L.J., Alvarez S., Phan H.T., Kondo Y., Kuriyan J, & Groves J.T. (2022). A two-component protein condensate of the EGFR cytoplasmic tail and Grb2 regulates Ras activation by SOS at the membrane. Proceedings of the National Academy of Sciences of the United States of America, 119(19), e2122531119.

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CREB Labeling with Alexa Fluors

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Exogenous and endogenous cysteines of CREB constructs were labeled with Alexa Fluor 488 and Alexa Fluor 647 maleimide dyes (Thermo Fisher). Purified CREB was labeled at 10–20 μM in labeling buffer (20 mM Tris pH 7.5, 500 mM NaCl, 1 mM TCEP) using a 5 mM dye stock. To avoid preferential labeling by one dye over the other, substoichiometric additions of the dye mixture were made to a purified protein construct over three hours to a final threefold molar excess of each dye. (For instance, six additions of dye mixture at half the concentration of protein were made every 30 minutes.) Labeled CREB was purified again by ion-exchange to remove excess fluorophores. bZIP constructs were labeled before cleavage of the GB1 tag for improved solubility. Purity of the labeled product was assessed via SDS-PAGE and mass spectrometry.

Li R.K., Ciblak M.A., Nordoff N., Pasarell L., Warnock D.W, & McGinnis M.R. (2000). In Vitro Activities of Voriconazole, Itraconazole, and Amphotericin B against Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum. Antimicrobial Agents and Chemotherapy, 44(6), 1734-1736.

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Lipid-based Single-Molecule Imaging

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Chemicals 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) (Ni 2+ -NTADOGS) were purchased from Avanti Polar Lipids. Alexa Fluor 488, Alexa Fluor 555, and Alexa Fluor 647 maleimide dyes were purchased from ThermoFisher. Bovine Serum Albumin (BSA), (±)-6-Hydroxy-2,5,7,8tetramethylchromane-2-carboxylic acid (Trolox), Catalase, 2-Mercaptoethanol (BME), NiCl2, H2SO4 and ATP were purchased from Sigma-Aldrich. Glucose Oxidase was purchased from Serva. Tris(2-carboxyethyl)phosphine (TCEP) was brought from Thermo Scientific. Glucose and H2O2 were from Fisher Scientific. MgCl2 was from EMD Chemicals. Tris buffer saline (TBS) was purchased from Corning.

Lin C., Nocka L.M., Stinger B., DeGrandchamp J., Lew N., Alvarez S., Phan H.T., Kondo Y., Kuriyan J., & Groves J.T. (2021). A two-component protein condensate of EGFR and Grb2 regulates Ras activation at the membrane.

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