ReadiLink™ Rapid XFD555 Antibody Labeling Kit XFD555 Same Structure to Alexa Fluor™ 555

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

Spectral properties

Correction Factor (260 nm)	0.08
Correction Factor (280 nm)	0.08
Extinction coefficient (cm ^-1 M ^-1)	150000
Excitation (nm)	553
Emission (nm)	568
Quantum yield	0.1¹

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™ Rapid iFluor® 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*

ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Production Scale*

Alternative formats

ReadiLink™ Rapid XFD555 Antibody Labeling Kit *Production Scale*

Overview SDS Protocol

Upgrade: ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 μg Antibody Per Reaction*, ReadiLink™ Rapid iFluor® 555 Antibody Labeling Kit *Production Scale*

Correction Factor (260 nm)

0.08

Correction Factor (280 nm)

0.08

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

150000

Excitation (nm)

553

Emission (nm)

568

Quantum yield

0.1¹

XFD555 is manufactured by AAT Bioquest, and it has the same chemical structure of Alexa Fluor® 555 (Alexa Fluor® is the trademark of ThermoFisher). Readilink™ protein labeling technology is the most robust and convenient tool for preparing fluorescent antibody conjugates used for fluorescence imaging and flow cytometry applications. ReadiLink™ Rapid XFD555 Antibody Labeling Kit provides the most convenient tool for making XFD555-labeled antibody conjugates. XFD555 dye used in this ReadiLink™ kit is reasonably stable and shows good reactivity and selectivity with protein amino groups. The kit has all the essential components for labeling ~2x50 ug antibody. Each of the two vials of XFD555 dye provided in the kit is optimized for labeling ~50 µg antibody. ReadiLink™ Rapid XFD555 Antibody Labeling Kit requires minimal hands-on time. The prepared XFD555 conjugates (with the kit) are ready to use for fluorescence imaging and flow cytometry applications without further purifications needed. It provides the most convenient method to prepare XFD555-labeled 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 ReadiUse™ Bio-Gel P-6 spin column (cat# 60500 from AAT Bioquest) 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) or gelatin will not 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-XFD555), 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

Correction Factor (260 nm)	0.08
Correction Factor (280 nm)	0.08
Extinction coefficient (cm ^-1 M ^-1)	150000
Excitation (nm)	553
Emission (nm)	568
Quantum yield	0.1¹

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (280 nm)	Correction Factor (260 nm)
ReadiLink™ Rapid FITC Antibody Labeling Kit Microscale Optimized for Labeling 50 μg Antibody Per Reaction	491	516	73000	0.92	0.35	-
ReadiLink™ Rapid Cy3 Antibody Labeling Kit Microscale Optimized for Labeling 50 μg Antibody Per Reaction	555	569	150000¹	0.15¹	0.073	0.07
ReadiLink™ Rapid Cy5 Antibody Labeling Kit Microscale Optimized for Labeling 50 μg Antibody Per Reaction	651	670	250000¹	0.27¹, 0.4²	0.03	0.02
ReadiLink™ Rapid Cy7 Antibody Labeling Kit Microscale Optimized for Labeling 50 μg Antibody Per Reaction	756	779	250000	0.3	0.036	0.05
ReadiLink™ Rapid XFD488 Antibody Labeling Kit XFD488 Same Structure to Alexa Fluor™ 488	499	520	71000	0.92¹	0.11	0.30
ReadiLink™ Rapid XFD594 Antibody Labeling Kit XFD594 Same Structure to Alexa Fluor™ 594	590	618	90000	0.66¹	0.56	0.43
ReadiLink™ Rapid XFD647 Antibody Labeling Kit XFD647 Same Structure to Alexa Fluor™ 647	650	671	239000	0.33¹	0.03	0.00
ReadiLink™ Rapid XFD750 Antibody Labeling Kit XFD750 Same Structure to Alexa Fluor™ 750	752	776	240000	0.12¹	0.04	0.00
ReadiLink™ Rapid FITC Antibody Labeling Kit Production Scale	491	516	73000	0.92	0.35	-
ReadiLink™ Rapid Cy3 Antibody Labeling Kit Production Scale	555	569	150000¹	0.15¹	0.073	0.07
ReadiLink™ Rapid Cy5 Antibody Labeling Kit Production Scale	651	670	250000¹	0.27¹, 0.4²	0.03	0.02
ReadiLink™ Rapid XFD488 Antibody Labeling Kit Production Scale	499	520	71000	0.92¹	0.11	0.30
ReadiLink™ Rapid XFD594 Antibody Labeling Kit Production Scale	590	618	90000	0.66¹	0.56	0.43
ReadiLink™ Rapid Cy7 Antibody Labeling Kit Production Scale	756	779	250000	0.3	0.036	0.05
ReadiLink™ Rapid XFD647 Antibody Labeling Kit Production Scale	650	671	239000	0.33¹	0.03	0.00
ReadiLink™ Rapid XFD750 Antibody Labeling Kit Production Scale	752	776	240000	0.12¹	0.04	0.00
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References

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Journal: Cellular and molecular neurobiology (2018): 171-180

Effective photo-enhancement of cellular activity of fluorophore-octaarginine antisense PNA conjugates correlates with singlet oxygen formation, endosomal escape and chromophore lipophilicity.
Authors: Yarani, Reza and Shiraishi, Takehiko and Nielsen, Peter E
Journal: Scientific reports (2018): 638

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