Antibody and Protein Labeling

Labeling an antibody with a specific tag has become an important and routine procedure in biological sciences and medical research. Tags, which range from visual markers such as fluorescent dyes, to hapten molecules like biotin, can be used to enhance the visibility of immune complexes and protein-protein interactions, or can be used in the isolation and purification of a protein.

When determining which type of label to choose focus on the downstream application for which the antibody will be used in. Fluorescent tags are used predominantly in cellular imaging, flow cytometry, fluorescent blotting and immunofluorescence applications, while enzymatic tags, such as HRP and AP, are commonly used for blotting, ELISAs and immunohistochemistry applications. Hapten tags such as biotin are the more versatile of the possible tag options as they can be used in protein isolation and purification, as well as, any of the aforementioned applications.

AAT Bioquest offers a comprehensive suite of antibody and protein labeling kits to choose from. Designed for the novice scientist in mind, these kits have the easiest protocols and require the least amount of hands-on experience.

Antibody Labeling Kits At-A-Glance

Antibody Labeling Kit ReadiLink™ Kits ReadiLink™ xtra Kits Buccutite™ Kits
Amount of antibody labeled per reaction 50 µg 50 µg 25 µg, 50 µg or 1 mg
Labeling Chemistry Targets free amine groups using amine-reactive dyes Targets free amine groups using amine-reactive dyes Uses two propietary linkers to facilitate protein-protein conjugation
Available labels iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7, Biotin iFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7 PE, APC, Tandem Dyes, HRP, AP
Type of bond between label and antibody Covalent Covalent Covalent
Compatible with BSA or other stabilizers? No Yes No
Requires purification? No No No
Time required for conjugation 1 hr 1 hr 2 hrs
Hands-on-time ∼10 minutes ∼10 minutes ∼15 minutes
Batch-to-batch variation Minimal Minimal Minimal
Applications¹ Imaging, FC, IF, WB Imaging, FC, IF, WB FC, IF, IHC, WB, ELISA
  1. FC = flow cytometry; IF = immunofluorescence; IHC = immunohistochemistry; WB = Western blot; ELISA = enzyme-linked immunosorbent assay.

 

ReadiLink™ Rapid Antibody Labeling Kits


AAT Bioquest ReadiLink™ Rapid Antibody Labeling Kits provide a quick and convenient method to label microscale volumes of antibodies with our superior iFluor™ dyes, mFluor™ dyes or other labels. The unique chemistry of ReadiLink™ kits enable researchers to effortlessly label and recover 100% of their antibodies without a purification step. Since ReadiLink™ conjugates are covalently labeled they are stable for long-term storage and they are ideal for demanding applications including fluorescent microscopy, multi-color imaging, multiplex flow cytometry, primary detection, IHC, FISH, ELISAs and more.

Key features of the ReadiLink™ Rapid Antibody Labeling Kits

  • Label monoclonal antibodies, polyclonal antibodies or other proteins
  • Quick and simple two-step labeling protocol, no chemistry or conjugation experience necessary
  • Useful for labeling 100 µg of antibody (two separate labeling reactions of 50 µg antibody)
  • 100% antibody recovery, NO purification of the product is required
  • Conjugates ready to use in 1 hr, with as little as ∼10 minutes hands-on time
  • Choice of iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5, Cy7, BSA, KLH or biotin
  • Avoid using any amine-containing buffers (e.g. Tris or glycine)
  • Stabilizing proteins (e.g. BSA or gelatin) must be removed prior to labeling, for antibody labeling kits compatible with BSA use ReadiLink™ xtra Rapid Antibody Labeling Kits

Convenient Design for Maximum Results

Figure 1. ReadiLink™ antibody labeling workflow (figure drawn in BioRender).

 

ReadiLink™ Rapid Antibody Labeling Kits utilize amine-reactive labels to prepare covalently labeled conjugates. These reactive labels selectively target and bind to primary amines, such as lysine residues, on the antibody of interest. To label your antibody, simply mix your antibody solution (concentration ∼1 mg/mL) with the ReadiLink™ reaction buffer, add the mixture to the vial of pre-formulated label, and incubate for 30 to 60 minutes. Lastly, add the TQ™-Dyed Quench Buffer to your mixture and mix well. This part is key as it eliminates both background interference and the column purification step. After 10 minutes of incubation your conjugates are ready to use.

 

High-performance alternative to Lightning-Link® Antibody Labeling Kits

Labeling Kit CD4(SK3)- Conjugate Stain Index
ReadiLink™ Kit CD4-iFluor™ 488 Conjugate 43
Lightning-Link® Kit CD4-Alexa Fluor® 488 Conjugate 26

 

Figure 2. Flow cytometry analysis of human PBMC cells. Anti-human CD4 antibody prepared with ReadiLink™ Rapid iFluor488 Antibody Labeling Kit (Cat No. 1255) or Lightning-Link® Rapid Alexa Fluor ® 488. The fluorescence signal was monitored using ACEA NovoCyte flow cytometer in the FITC channel.

Table 1. ReadiLink™ Rapid Antibody Labeling Kit Selection Guide

Fluorophore Fluorophore Equivalent Ex/Em (nm) Imaging Flow Cytometry Cat No.
mFluor™ Violet 420 Cascade® Blue 398/411  – Yes 1105
iFluor™ 350 AMCA, Alexa Fluor® 350 345/442 Yes  – 1220
mFluor™ Violet 450 Pacific Blue® 403/454  – Yes 1100
mFluor™ Violet 510 –  414/508  – Yes 1110
iFluor™ 488 FITC, Alexa Fluor® 488 491/514 Yes Yes 1255
FITC –  494/520 Yes Yes 1299
mFluor™ Violet 540 Pacific Orange® 405/537  – Yes 1114
Cy3 –  555/565 Yes  – 1290
iFluor™ 555 Cy3, Alexa Fluor® 555 552/567 Yes  – 1227
mFluor™ Blue 570 R-PE 553/570  – Yes 1120
iFluor™ 594 Texas Red®, Alexa Fluor® 594 592/614 Yes –  1230
mFluor™ Green 620  – 522/617  – Yes 1123
iFluor™ 633 Cy5, Alexa Fluor® 633 638/655 Yes Yes 1260
Cy5  – 644/665 Yes Yes 1292
iFluor™ 647 Cy5, Alexa Fluor® 647 654/674 Yes Yes 1235
mFluor™ Red 700 APC-Alexa Fluor® 700 Tandem 657/700 Yes 1130
iFluor™ 680 Cy5.5, Alexa Fluor® 680, IRDye® 700 682/701 Yes Yes 1240
iFluor™ 700 Alexa Fluor® 700 693/713 Yes Yes 1245
Cy7  – 749/776 Yes  – 1294
iFluor™ 750 Cy7, Alexa Fluor® 750 753/779 Yes  – 1250
mFluor™ Red 780 APC-Alexa Fluor® 750 Tandem 629/780  – Yes 1131
iFluor™ 790 IRDye® 800, Alexa Fluor® 790 782/811 Yes  – 1265

BSA Compatible ReadiLink™ xtra Rapid Antibody Labeling Kits


ReadiLink™ xtra Rapid Antibody Labeling Kits with BSA compatibility takes labeling primary antibodies to a whole new level of convenience. Utilizing an optimized labeling protocol combined with vials of specially formulated labels, ReadiLink™ xtra kits enables rapid and reproducible labeling of microgram quantites of a primary antibody in the presence of common buffer additives and stabilizing proteins. In just two simple steps and without column purification, covalently label antibodies that are suitable for use in any immuno-based application.

Key features of the ReadiLink™ xtra Rapid Antibody Labeling Kits

  • Compatible with BSA and other stabilizing proteins
  • Label monoclonal antibodies, polyclonal antibodies or other proteins
  • Labels 100 µg of antibody (two separate labeling reactions of 50 µg antibody)
  • 100% antibody recovery, NO purification of the product is required
  • Conjugates ready to use in 1 hr, with as little as ∼10 minutes hands-on time
  • Choice of iFluor™ dyes, mFluor™ dyes, Alexa Fluor® equivalent AF dyes, FITC, Cy3, Cy5 or Cy7

 

 

Table 2. ReadiLink™ xtra Rapid Antibody Labeling Kits

Name Ex (nm) Em (nm) Unit Size Cat No.
ReadiLink™ xtra Rapid iFluor™ 350 Antibody Labeling Kit 344 448 2 Labelings 1950
ReadiLink™ xtra Rapid iFluor™ 488 Antibody Labeling Kit 491 516 2 Labelings 1955
ReadiLink™ xtra Rapid AF488 Antibody Labeling Kit 499 520 2 Labelings 1978
ReadiLink™ xtra Rapid FITC Antibody Labeling Kit 491 516 2 Labelings 1970
ReadiLink™ xtra Rapid iFluor™ 555 Antibody Labeling Kit 556 569 2 Labelings 1958
ReadiLink™ xtra Rapid AF555 Antibody Labeling Kit 553 568 2 Labelings 1980
ReadiLink™ xtra Rapid iFluor™ 594 Antibody Labeling Kit 587 603 2 Labelings 1960
ReadiLink™ xtra Rapid AF594 Antibody Labeling Kit 590 618 2 Labelings 1982
ReadiLink™ xtra Rapid iFluor™ 647 Antibody Labeling Kit 654 669 2 Labelings 1963
ReadiLink™ xtra Rapid AF647 Antibody Labeling Kit 650 671 2 Labelings 1985
ReadiLink™ xtra Rapid iFluor™ 750 Antibody Labeling Kit 759 777 2 Labelings 1965
ReadiLink™ xtra Rapid AF750 Antibody Labeling Kit 752 776 2 Labelings 1988

Buccutite™ Protein to Protein Labeling Kits


Buccutite™ crosslinking technology offers a trouble-free way to covalently label proteins and antibodies with horseradish peroxidase (HRP), alkaline phosphatase (AP) or a phycobiliprotein. Compared to the conventional techniques for protein-protein conjugation, such as the SMCC chemical labeling method, Buccutite™ conjugation is significantly more robust and less demanding. These kits allow optimal labeling of microgram amounts of antibodies (as little as 25 µg per labeling reaction) in less than two hours, with no purification necessary and 100% yield. With Buccutite™ labeling kits, routinely prepare covalently-labeled conjugates that will withstand the rigorous washing techniques generally required in a typical immunoassay (e.g. ELISA, IHC and Western blot) or flow cytometry application.

Key features of the Buccutite™ Rapid Antibody Labeling Kit

  • Label 25 µg up to 1 mg of antibody
  • Choice of labels include HRP, AP, phycoerythrin (PE), allophycocyanin (APC), PerCP, PE-tandem dyes and APC tandem dyes
  • Less than 2 hours of labeling time, with no purification required
  • 100% antibody recovery

 

High-performance alternative to Lightning-Link® Antibody Labeling Kits

Labeling Kit CD4(SK3)- Conjugate Stain Index
Buccutite™ Kit CD4-PE Conjugate 55
Lightning-Link® Kit CD4-PE Conjugate 28

 

Figure 3. Flow cytometry analysis of human PBMC cells. Anti-human CD4 antibody prepared with Buccutite™ Rapid PE Antibody Labeling Kit (Cat No. 1310) or Lightning-Link® PE. The fluorescence signal was monitored using ACEA NovoCyte flow cytometer in the FITC channel.

Table 3. Buccutite™ Rapid Antibody Labeling Kits

Name Ex (nm) Em (nm) Unit Size Cat No.
Buccutite™ Rapid Protein Crosslinking Kit *Microscale Optimized for Crosslinking 100 ug Antibody Per Reaction* 2 Labelings 1315
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 1 mg Protein* 1 Kit 5504
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 100 ug Protein* 1 Kit 5503
Buccutite™ Peroxidase (HRP) Antibody Conjugation Kit *Optimized for Labeling 25 ug Protein* 2 Labelings 5505
Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 565 575 2 Labelings 1310
Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 565 575 2 Labelings 1312
Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 565 600 2 Labelings 1318
Buccutite™ Rapid PE-Texas Red Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 565 600 2 Labelings 1343
Buccutite™ Rapid PE-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 565 674 2 Labelings 1322
Buccutite™ Rapid PE-Cy5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 565 674 2 Labelings 1340
Buccutite™ Rapid PE-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 565 700 2 Labelings 1316
Buccutite™ Rapid PE-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 565 700 2 Labelings 1341
Buccutite™ Rapid PE-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 565 780 2 Labelings 1317
Buccutite™ Rapid PE-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 565 780 2 Labelings 1342
Buccutite™ Rapid PerCP Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 482 677 2 Labelings 1325
Buccutite™ Rapid PerCP Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 482 677 2 Labelings 1353
Buccutite™ Rapid APC Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 651 665 2 Labelings 1311
Buccutite™ Rapid APC Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 651 665 2 Labelings 1313
Buccutite™ Rapid APC-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 651 700 2 Labelings 1320
Buccutite™ Rapid APC-Cy5.5 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 651 700 2 Labelings 1350
Buccutite™ Rapid APC-iFluor™ 700 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 651 713 2 Labelings 1319
Buccutite™ Rapid APC-iFluor™ 700 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 651 713 2 Labelings 1347
Buccutite™ Rapid APC-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction* 651 780 2 Labelings 1321
Buccutite™ Rapid APC-Cy7 Tandem Antibody Labeling Kit *Microscale Optimized for Labeling 25 ug Antibody Per Reaction* 651 780 2 Labelings 1351

Product Ordering Information


 

Table 4. Ordering Info for Readilink Antibody Labeling Kits Products

Cat# Product Name Unit Size Ex Em
1100 ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 403 454
1105 ReadiLink™ Rapid mFluor™ Violet 420 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 398 411
1110 ReadiLink™ Rapid mFluor™ Violet 510 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 414 508
1114 ReadiLink™ Rapid mFluor™ Violet 540 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 405 537
1120 ReadiLink™ Rapid mFluor™ Blue 570 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 553 570
1227 ReadiLink™ Rapid iFluor™ 555 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 552 567
1230 ReadiLink™ Rapid iFluor™ 594 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 592 614
1235 ReadiLink™ Rapid iFluor™ 647 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 654 674
1240 ReadiLink™ Rapid iFluor™ 680 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 682 701
1245 ReadiLink™ Rapid iFluor™ 700 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 693 713
1250 ReadiLink™ Rapid iFluor™ 750 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 753 779
1255 ReadiLink™ Rapid iFluor™ 488 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 491 514
1260 ReadiLink™ Rapid iFluor™ 633 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 638 655
1265 ReadiLink™ Rapid iFluor™ 790 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 782 811
1290 ReadiLink™ Rapid Cy3 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 555 565
1292 ReadiLink™ Rapid Cy5 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 649 665
1294 ReadiLink™ Rapid Cy7 Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 749 776
1299 ReadiLink™ Rapid FITC Antibody Labeling Kit *Microscale Optimized for Labeling 50 µg Antibody Per Reaction* 2 Labelings 492 516
1300 ReadiLink™ Rapid trFluor™ Eu Antibody Labeling Kit *Microscale Optimized for Labeling 50 ug Antibody Per Reaction* 2 Labelings 346 617
5400 ReadiLink™ Protein Conjugation Stop Buffer 1 ml
5501 ReadiLink™ BSA Conjugation Kit 1 kit
5502 ReadiLink™ KLH Conjugation Kit 1 kit
5520 ReadiLink™ Protein Biotinylation Kit 2 Labelings
5521 ReadiLink™ Protein Biotinylation Kit *Powered by ReadiView™ Biotin Visionization Technology* 1 kit

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