ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction*

! Interactive Product Finder: This product has alternative forms and/or upgrades. View now


Image Viewer
Select color channel(s)
HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by AAT’s Cy5<sup>®</sup> goat anti-mouse IgG conjugate (Red, Left) or Jackson’s Cy5<sup>®</sup> goat anti-mouse IgG conjugate (Red, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue, Cat# 17530).
Roll over image to zoom in


Loading...
 
Unit Size: Cat No: Price (USD): Qty:
2 Labelings 1292 $145


Export item/cart as Excel file

Send item/cart as email
EXPORT TO EXCEL X

Export:
EXPORT TO EMAIL X
Important: We request your email address to ensure that the recipient(s) knows you intended for them to see the email, and that it is not junk mail.
Export:
Your Name*:
Your Email*:
Recipient Email*:
Your Personal Message:
Additional Ordering Information
Telephone: 1-800-990-8053
Fax: 1-408-733-1304
Email: sales@aatbio.com
International: See distributors





Overview

SolventDMSO
Storage Freeze (<-15 °C)
Minimize light exposure
Category Superior Labeling Dyes
iFluor Dyes and Kits
Related General proteins
Labeling via Amino Groups
Cy5 is one of the most popular fluorescent labeling dyes for preparing orange-red fluorescent bioconjugates. However, most of the commercial Cy5 labeling kits require intensive hands-on time. This Cy5 ReadiLink™ labeling kit is one of the most robust protein labeling kits for preparing Cy5-labeled antibody conjugates or other protein conjugates. It essentially only requires 2 simple mixing steps without a column purification required. The kit provides all the essential components for labeling ~2x50 ug antibody. Each of the two vials of Cy5 dye provided in the kit is optimized for labeling ~50 µg antibody. This Cy5 protein labeling kit provides a convenient method to label monoclonal, polyclonal antibodies or other proteins (>10 kDa).




Spectrum Advanced Spectrum Viewer

Sorry, your browser does not support inline SVG. Relative Intensity (%) 100 80 60 40 20 0 Sorry, your browser does not support inline SVG.
Sorry, your browser does not support inline SVG. Sorry, your browser does not support inline SVG.
Move mouse over grid to display wavelength & intensity values.

300
400
500
600
700
800
900
Wavelength (nm)





Protocol


Quick Preview

This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

Table 1. Available fluorophores in AAT Bioquest ReadyLink™ Rapid Antibody Labelling Kits

Cat# Labels Ex (nm) Em (nm)
1100  mFluor™ Violet 450 403 454
1105 mFluor™ Violet 420 398 411
1110 mFluor™ Violet 510  414  508
1114 mFluor™ Violet 540  399  550
1120 mFluor™ Blue 570 553  570 
1123 mFluor™ Green 620  522  617
1126 mFluor™ Yellow 630  561  630
1130 mFluor™ Red 700   657 700 
1131  mFluor™ Red 780 629  780 
1220 iFluor™ 350   345 442
1227 iFluor™ 555  559  569
1230  iFluor™ 594  592 614
1235  iFluor™ 647  654 674
1240  iFluor™ 680  682 701
1245  iFluor™ 700  693 713
1250  iFluor™ 750  753 779
1255  iFluor™ 488  491 514
1260  iFluor™ 633  638 655
1265 iFluor™ 790   782 811
1290  Cy3  555 565
1292  Cy5  644 665
1294  Cy7  749  776
1299  FITC 494  520
Preparation of working solution

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.

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) 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 

  1. 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.

  2. 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

  1. 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.

  2. 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.

Example data analysis and figures

Figure 1. HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by AAT’s Cy5® goat anti-mouse IgG conjugate (Red, Left) or Jackson’s Cy5® goat anti-mouse IgG conjugate (Red, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue, Cat# 17530).
Disclaimer
AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email info@aatbio.com if you have any questions.









References

Cube-shaped theranostic paclitaxel prodrug nanocrystals with surface functionalization of SPC and MPEG-DSPE for imaging and chemotherapy
Authors: Fuqiang Guo, Jiajia Shang, Hai Zhao, Kangrong Lai, Yang Li, Zhongxiong Fan, Zhenqing Hou, Guanghao Su
Journal: Colloids and Surfaces B: Biointerfaces (2017)

Light/magnetic hyperthermia triggered drug released from multi-functional thermo-sensitive magnetoliposomes for precise cancer synergetic theranostics
Authors: Yuxin Guo, Yang Zhang, Jinyuan Ma, Qi Li, Yang Li, Xinyi Zhou, Dan Zhao, Hua Song, Qing Chen, Xuan Zhu
Journal: Journal of Controlled Release (2017)

Thermo-sensitive hydrogel PLGA-PEG-PLGA as a vaccine delivery system for intramuscular immunization
Authors: Xiaoyan Wang, Yu Zhang, Wei Xue, Hong Wang, Xiaozhong Qiu, Zonghua Liu
Journal: Journal of Biomaterials Applications (2017): 923--932

Affinity-Controlled Protein Encapsulation into Sub-30 nm Telodendrimer Nanocarriers by Multivalent and Synergistic Interactions
Authors: Xu Wang, Changying Shi, Li Zhang, Alexa Bodman, Dandan Guo, Lili Wang, Walter A Hall, Stephan Wilkens, Juntao Luo
Journal: Biomaterials (2016)

Carboxymethyl Dextran-Stabilized Polyethylenimine-Poly (epsilon-caprolactone) Nanoparticles-Mediated Modulation of MicroRNA-34a Expression via Small-Molecule Modulator for Hepatocellular Carcinoma Therapy
Authors: Xiongwei Deng, Zhaoxia Yin, Zhixiang Zhou, Yihui Wang, Fang Zhang, Qin Hu, Yishu Yang, Jianqing Lu, Yan Wu, Wang Sheng
Journal: ACS applied materials & interfaces (2016): 17068--17079

Click-electron microscopy for imaging metabolically tagged nonprotein biomolecules
Authors: John T Ngo, Stephen R Adams, Thomas J Deerinck, Daniela Boassa, Frances Rodriguez-Rivera, Sakina F Palida, Carolyn R Bertozzi, Mark H Ellisman, Roger Y Tsien
Journal: Nat Chem Biol (2016): 459--465

Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery
Authors: Wen Zhao, Yifan Zhang, Xueyun Jiang, Chunying Cui
Journal: Drug Design, Development and Therapy (2016): 3851

Molecular Basis and Consequences of the Cytochrome c-tRNA Interaction
Authors: Cuiping Liu, Aaron J Stonestrom, Thomas Christian, Jeongsik Yong, Ryuichi Takase, Ya-Ming Hou, Xiaolu Yang
Journal: Journal of Biological Chemistry (2016): 10426--10436

Determination of the active transport of fucoidan derived from okinawa mozuku across the human intestinal caco-2 cells as assessed by size-exclusion chromatography
Authors: Takeaki Nagamine, Kou Hayakawa, Kyoumi Nakazato, Masahiko Iha
Journal: Journal of Chromatography B (2015): 187--193

Multiplexed single-cell in situ RNA analysis by reiterative hybridization
Authors: Lu Xiao, Jia Guo
Journal: Analytical Methods (2015): 7290--7295


View More Citations




Resources
 
Safety Data Sheet (SDS)


Documents
1. Fluorescent Labeling Probes & Kits

Certificate of Analysis