Antibody Conjugation Kits

ReadiLink™ Antibody Conjugation Kits

Antibody conjugation in a range of fluorophores

Use ReadiLink Antibody Conjugation Kits to quickly conjugate microscale volumes of antibody (50-100 µg) in two easy steps with the unique chemistry of ReadiLink Dyes. Fluorophores are available that are excited by UV all the way to infrared with a range of emission spectra to easily fit into your multicolor flow cytometry experiments, giving you greater choice.

Figure 1. Efficient labeling of antibodies in two easy steps

Fig. 1. Efficient labeling of antibodies in two easy steps

The unique chemistry of ReadiLink Dyes allows you to quickly label your antibody in just two easy steps, shown in Figure 1.

Each ReadiLink Dye Kit includes succinimidyl esters that selectively react with primary amines to form a stable carboxamide bond. This bond ensures no dissociation between the fluorophore and antibody which may lead to unbound fluorophore in your experiments.


ReadiLink Antibody Conjugation Kits key benefits:
 

  • Saving you time: As no purification column required

  • Minimal hands on time: Easy protocol doesn’t involve complicated chemistry, allowing you to perform other experiments

  • Ready-to-use: No fluorophore quenching required

  • Versatile: Available in 13 unique fluorophores with a wide range of excitation wavelengths – giving you greater flexibility in your multicolor flow cytometry and cell sorting experiments as well as other applications such as western blot and ELISA

ReadiLink Conjugation Kits allow you to label your primary antibody quickly avoiding the use of secondary antibodies and can help improve your flexibility in building panels. Purified CD45RA was labeled with ReadiLink 405/454 1351011 and human peripheral blood stained in combination with CD3 FITC (MCA463F) and CD4 (MCA1267PE). When compared to a Pacific Blue™ directly conjugated CD45RA (MCA88PB)  antibody the staining was comparable allowing identification of naïve CD4 positive T cells.

Fig 1. Purified CD45RA (MCA88) was labeled with ReadiLink 405/454 and used to stain human peripheral blood in combination with CD4 PE (MCA1267PE).

Fig.2. Purified CD45RA (MCA88) was labeled with ReadiLink 405/454 and used to stain human peripheral blood in combination with CD4 PE (MCA1267PE). Alternatively directly labeled CD45RA Pacific Blue (MCA88PB) was used. The staining shown is lymphocytes gated on the CD3 positive population. CD4 and CD45RA positive naïve T cells can be identified in both plots.

Fig 2. Purified CD56 (MCA2693) was labeled with ReadiLink 405/537 and used to stain human peripheral blood in combination with CD3 FITC (MCA463F).

Fig.3. Purified CD56 (MCA2693) was labeled with ReadiLink 405/537 and used to stain human peripheral blood in combination with CD3 FITC (MCA463F).

The staining shown is lymphocytes gated on the CD3 positive population. CD56 positive NK cells can be clearly identified (red circle).

Using our ReadiLink kits even low expressing antigens can be identified. CD56 (MCA2693) was conjugated to ReadiLink 405/537 1351013 and counterstained with CD3 FITC (MCA463F). As can be seen in Figure 3, NK cells can be clearly defined.

The table below shows the range of available ReadiLink Dyes, maximal excitation and emission properties, and their fluorophore equivalents to enable you to choose the right fluorophore for your flow cytometry experiment.

ReadiLink Antibody Labeling Kits Fluorophore Equivalents Maximum Excitation, nm Maximum Emission, nm S3™ Cell Sorter Laser, nm 488/561 nm Filter Channel Catalog Number
ReadiLink 
350/440

AMCA
AlexaFluor®:350
DyLight 350

350 440 1351001
ReadiLink 
405/454
Pacific Blue 403 454 1351011
ReadiLink
405/508
414 508 1351012
ReadiLink 
405/537
Pacific Orange 405 537 1351013
ReadiLink 
492/516
FITC
AlexaFluor 488
DyLight 488
492 516 488 FL1
(525/30)
1351002
ReadiLink 
555/570
Cy3
AlexaFluor 555
DyLight 550
555 570 561 FL2
(586/25)
1351003
ReadiLink 
594/610

Texas Red
AlexaFluor 594
DyLight 594

594 610 561 FL3
(615/25)
1351004
ReadiLink
633/655
Cy5
AlexaFluor 633
DyLight 633
633 655 1351005
ReadiLink 
647/674
Cy5
AlexaFluor 647
DyLight 650
647 674 1351006
ReadiLink
680/701
Cy5.5
IRDye 700
AlexaFluor 680
DyLight 680
680 701 1351007
ReadiLink 
700/713
AlexaFluor 700 700 713 1351008
ReadiLink
750/780
AlexaFluor 750
DyLight 755
750 780 1351009
ReadiLink
790/811
IRDye 800
AlexaFluor 790
DyLight 800
790 811 1351010

In addition to conjugation kits we have flow cytometry validated antibodies, reagents and resources to help you optimize your flow cytometry.­­­



ReadiLink™ is a trademark of AAT Bioquest, Inc.
Alexa Fluor®: Molecular Probes, Inc, OR, USA
Cy containing products or portions thereof are manufactured under license from Carnegie Mellon University under U.S. Patent Number 5,268,486 and related patents
Cy® is a trademark of GE Healthcare Limited
Pacific Blue™ is a trademark of Molecular Probes Inc, OR, USA