Aladdin Fluo™ 647 C₂-maleimide - Ex:651nm, Em:670nm , CAS No.A638836

CAS: A638836 Cat. No.: A638836

GRADE & PURITY Em:670nm, Ex:651nm

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Descripción general

Aladdin Fluo 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, Aladdin Fluo 647 dye is water soluble and pH-insensitive from pH 4 to pH 10. In addition to reactive dye formulations, we offer Aladdin Fluo 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 Aladdin Fluo 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 Aladdin Fluo 647 conjugates exhibit brighter fluorescence and greater photostability than the conjugates of other spectrally similar fluorophores.
Detailed information about this Aladdin Fluo maleimide:
Fluorophore label: Aladdin Fluo 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 Aladdin Fluo 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 Aladdin Fluo dye using a gel filtration column. 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 ()

Specifications

Especificaciones y pureza

Ex:651nm, Em:670nm

Condiciones de almacenamiento de almacenamiento

Protected from light, Store at -20°C

Enviado en

Ice chest + Ice pads

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Grado

Em:670nm, Ex:651nm

Propiedades del producto

• Reactividad: Thiol

Propiedades químicas y físicas

• Sensibilidad: light sensitive