ReadiLink™ xtra Rapid XFD647 Antibody Labeling Kit BSA-Compatible, XFD647 Same Structure to Alexa Fluor™ 647

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
Quotation	Request
International	See distributors
Shipping	Standard overnight for United States, inquire for international

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	270000
Excitation (nm)	650
Emission (nm)	671
Quantum yield	0.33¹

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12171501

Direct upgrades

ReadiLink™ xtra Rapid iFluor® 647 Antibody Labeling Kit *BSA-Compatible*

Overview SDS Protocol

Upgrade: ReadiLink™ xtra Rapid iFluor® 647 Antibody Labeling Kit *BSA-Compatible*

Extinction coefficient (cm ^-1 M ^-1)

270000

Excitation (nm)

650

Emission (nm)

671

Quantum yield

0.33¹

XFD647 is manufactured by AAT Bioquest, and it has the same chemical structure of Alexa Fluor® 647 (Alexa Fluor® is the trademark of ThermoFisher). ReadiLink™ xtra rapid antibody labeling kits require essentially only 2 simple mixing steps without a column purification needed. Specially formulated and preactivated XFD647 (chemically equivalent to Alexa Fluor® 647) used in this ReadiLink™ kit is quite stable and shows good reactivity and selectivity with antibodies. The kit has all the essential components for labeling ~2x50 ug antibody. Each of the two vials of preactivated XFD647 dye provided in the kit is optimized for labeling ~50 µg antibody. ReadiLink™ xtra XFD647 rapid antibody labeling kit provides a convenient and robust method to label monoclonal and polyclonal antibodies with red fluorescent XFD647 fluorophore. XFD647 is one of the most used fluorophores for labeling antibodies.

Components

Example protocol

AT A GLANCE

Important

Warm all the components and centrifuge the vials briefly before opening, and immediately prepare the required solutions before starting your conjugation. The following protocol is for recommendation.

PREPARATION OF WORKING SOLUTION

Protein working solution (Solution A)

For labeling 50 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 5 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 50 µL of the target protein solution. Note: If you have a different protein concentration, adjust the protein volume accordingly to make ~50 µg of protein available for your labeling reaction. Note: For labeling 100 µg of protein (assuming the target protein concentration is 1 mg/mL), mix 10 µL (10% of the total reaction volume) of Reaction Buffer (Component B) with 100 µL of the target protein solution. Note: The protein should be dissolved in 1X phosphate buffered saline (PBS), pH 7.2 - 7.4; if the protein is dissolved in glycine buffer, it must be dialyzed against 1X PBS, pH 7.2 - 7.4, or use Amicon Ultra-0.5, Ultracel-10 Membrane, 10 kDa (cat# UFC501008 from Millipore) to remove free amines or ammonium salts (such as ammonium sulfate and ammonium acetate) that are widely used for protein precipitation. Note: Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) with 0.1 to 0.5 % will be labeled well. Note: For optimal labeling efficiency, a final protein concentration range of 1 - 2 mg/mL is recommended, with a significantly reduced conjugation efficiency at less than 1 mg/mL.

SAMPLE EXPERIMENTAL PROTOCOL

Run conjugation reaction

Add the protein working solution (Solution A) to ONE vial of labeling dye (Component A), and mix them well by repeatedly pipetting for a few times or vortex the vial for a few seconds. Note: If labeling 100 µg of protein, use both vials (Component A) of labeling dye by dividing the 100 µg of protein into 2 x 50 µg of protein and reacting each 50 µg of protein with one vial of labeling dye. Then combine both vials for the next step.
Keep the conjugation reaction mixture at room temperature for 30 - 60 minutes. Note: The conjugation reaction mixture can be rotated or shaken for longer time if desired.

Stop Conjugation reaction

Add 5 µL (for 50 µg protein) or 10 µL (for 100 µg protein) which is 10% of the total reaction volume of TQ™-Dyed Quench Buffer (Component C) into the conjugation reaction mixture; mix well.
Incubate at room temperature for 10 minutes. The labeled protein (antibody) is now ready to use.

Storage of Protein Conjugate

The protein conjugate should be stored at > 0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin). For longer storage, the protein conjugates could be lyophilized or divided into single-used aliquots and stored at ≤ –20°C.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	270000
Excitation (nm)	650
Emission (nm)	671
Quantum yield	0.33¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (280 nm)
ReadiLink™ xtra Rapid FITC Antibody Labeling Kit BSA-Compatible	491	516	73000	0.92	0.35
ReadiLink™ xtra Rapid Cy3 Antibody Labeling Kit BSA-Compatible	555	569	150000¹	0.15¹	0.073
ReadiLink™ xtra Rapid Cy5 Antibody Labeling Kit BSA-Compatible	651	670	250000¹	0.27¹, 0.4²	0.03
ReadiLink™ xtra Rapid Cy7 Antibody Labeling Kit BSA-Compatible	756	779	250000	0.3	0.036
ReadiLink™ xtra Rapid XFD594 Antibody Labeling Kit BSA-Compatible, XFD594 Same Structure to Alexa Fluor™ 594	590	618	92000	0.66¹	-
ReadiLink™ xtra Rapid XFD488 Antibody Labeling Kit BSA-Compatible, XFD488 Same Structure to Alexa Fluor™ 488	499	520	73000	0.92¹	-
ReadiLink™ xtra Rapid XFD555 Antibody Labeling Kit BSA-Compatible, XFD555 Same Structure to Alexa Fluor™ 555	553	568	155000	0.1¹	-
ReadiLink™ xtra Rapid XFD750 Antibody Labeling Kit BSA-Compatible, XFD750 Same Structure to Alexa Fluor™ 750	752	776	290000	0.12¹	-

References

View all 50 references: Citation Explorer

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Authors: Ovejero Paredes, Karina and Díaz-García, Diana and García-Almodóvar, Victoria and Lozano Chamizo, Laura and Marciello, Marzia and Díaz-Sánchez, Miguel and Prashar, Sanjiv and Gómez-Ruiz, Santiago and Filice, Marco
Journal: Cancers (2020)

Visualization of Mineral-Targeted Alkaline Phosphatase Binding to Sites of Calcification In Vivo.
Authors: Amadeu de Oliveira, Flavia and Narisawa, Sonoko and Bottini, Massimo and Millán, José Luis
Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research (2020)

Long-Range Single-Molecule Förster Resonance Energy Transfer between Alexa Dyes in Zero-Mode Waveguides.
Authors: Baibakov, Mikhail and Patra, Satyajit and Claude, Jean-Benoît and Wenger, Jérôme
Journal: ACS omega (2020): 6947-6955

Subtype-Selective Fluorescent Ligands as Pharmacological Research Tools for the Human Adenosine A2A Receptor.
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Journal: Journal of medicinal chemistry (2020): 2656-2672

Electrostatically gated nanofluidic membrane for ultra-low power controlled drug delivery.
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Effect of Vectashield-induced fluorescence quenching on conventional and super-resolution microscopy.
Authors: Arsić, Aleksandra and Stajković, Nevena and Spiegel, Rainer and Nikić-Spiegel, Ivana
Journal: Scientific reports (2020): 6441

Sequential energy transfer driven by monoexponential dynamics in a biohybrid light-harvesting complex 2 (LH2).
Authors: Yoneda, Yusuke and Kato, Daiji and Kondo, Masaharu and Nagashima, Kenji V P and Miyasaka, Hiroshi and Nagasawa, Yutaka and Dewa, Takehisa
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Detection of the Entry of Nonlabeled Transportan 10 into Single Vesicles.
Authors: Shuma, Madhabi Lata and Moghal, Md Mizanur Rahman and Yamazaki, Masahito
Journal: Biochemistry (2020)

Photo-isomerization of the Cyanine Dye Alexa-Fluor 647 (AF-647) in the Context of dSTORM Super-Resolution Microscopy.
Authors: Karlsson, Joshua K G and Laude, Alex and Hall, Michael J and Harriman, Anthony
Journal: Chemistry (Weinheim an der Bergstrasse, Germany) (2019): 14983-14998

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