PE/Cy7® Conjugation Kit is a simple and quick process for antibody labeling/conjugation
- Rapid Conjugation: PE/Cy7® labeling in <4 hrs with just 30 secs of hands-on time.
- High Efficiency: Ensures 100% antibody recovery, preserving the integrity and value of your antibodies.
- Versatility: Suitable for conjugating antibodies, proteins, and peptides. The labeled antibodies can be used immediately in applications such as Western Blot, ELISA, and IHC without further purification.
- Confidence: Cited in over 15 publications.
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- The Journal of clinical investigation 134:2024Inhibiting the NADase CD38 improves cytomegalovirus-specific CD8+ T cell functionality and metabolism.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNils Mülling,Felix M Behr,Graham A Heieis,Kristina Boss,Suzanne van Duikeren,Floortje J van Haften,Iris N Pardieck,Esmé Ti van der Gracht,Ward Vleeshouwers,Tetje C van der Sluis,J Fréderique de Graaf,Dominique Mb Veerkamp,Kees Lmc Franken,Xin Lei,Lukas van de Sand,Sjoerd H van der Burg,Marij Jp Welters,Sebastiaan Heidt,Wesley Huisman,Simon P Jochems,Martin Giera,Oliver Witzke,Aiko Pj de Vries,Andreas Kribben,Bart Everts,Benjamin Wilde,Ramon ArensPubMed 38954588
- Nature communications 14:70852023Tunable Tamm plasmon cavity as a scalable biosensing platform for surface enhanced resonance Raman spectroscopy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKandammathe Valiyaveedu Sreekanth,Jayakumar Perumal,U S Dinish,Patinharekandy Prabhathan,Yuanda Liu,Ranjan Singh,Malini Olivo,Jinghua TengPubMed 37925522
- Nature communications 14:56272023Metabolic heterogeneity of tissue-resident macrophages in homeostasis and during helminth infection.Applications:Flow Cyt (Intra)Reactive species:MouseGraham A Heieis,Thiago A Patente,Luís Almeida,Frank Vrieling,Tamar Tak,Georgia Perona-Wright,Rick M Maizels,Rinke Stienstra,Bart EvertsPubMed 37699869
- Current protocols 2:e5842022Evaluating the Clinical and Immune Responses to Spotted Fever Rickettsioses in the Guinea Pig-Tick-Rickettsia System.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJohn V Stokes,Michael L Levin,Claire E Cross,Anne-Marie L Ross,Alyssa N Snellgrove,Bridget V Willeford,Navatha Alugubelly,Andrea S Varela-StokesPubMed 36383032
- iScience 25:1043842022Contrasting behavior between the three human monocyte subsets in dengue pathophysiology.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDeepti Maheshwari,Keshav Saini,Prabhat Singh,Mohit Singla,Kaustuv Nayak,Charu Aggarwal,Yadya M Chawla,Prashant Bajpai,Manpreet Kaur,Sivaram Gunisetty,Christiane S Eberhardt,Rajni Nyodu,Kathryn Moore,Mehul S Suthar,Guruprasad R Medigeshi,Evan Anderson,Rakesh Lodha,Sushil K Kabra,Rafi Ahmed,Anmol Chandele,Kaja Murali-KrishnaPubMed 35620424
- iScience 24:1033122021Reduced mitochondrial respiration in T cells of patients with major depressive disorder.Applications:Unspecified applicationReactive species:Unspecified reactive speciesStefanie Gamradt,Helge Hasselmann,Aline Taenzer,Jelena Brasanac,Victoria Stiglbauer,Arne Sattler,Max Sajitz-Hermstein,Sylwia Kierszniowska,Caren Ramien,Jan Nowacki,Lea Mascarell-Maricic,Katja Wingenfeld,Dominique Piber,Andreas Ströhle,Katja Kotsch,Friedemann Paul,Christian Otte,Stefan M GoldPubMed 34765928
- Molecular oncology 15:2330-23442021Development and initial clinical testing of a multiplexed circulating tumor cell assay in patients with clear cell renal cell carcinoma.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRory M Bade,Jennifer L Schehr,Hamid Emamekhoo,Benjamin K Gibbs,Tamara S Rodems,Matthew C Mannino,Joshua A Desotelle,Erika Heninger,Charlotte N Stahlfeld,Jamie M Sperger,Anupama Singh,Serena K Wolfe,David J Niles,Waddah Arafat,John A Steinharter,E Jason Abel,David J Beebe,Xiao X Wei,Rana R McKay,Toni K Choueri,Joshua M LangPubMed 33604999
- Scientific reports 11:10632021Tick saliva-induced programmed death-1 and PD-ligand 1 and its related host immunosuppression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYamato Sajiki et. al.PubMed 33441793
- Cellular and molecular gastroenterology and hepatology 11:1045-10692020Cholangiopathy and Biliary Fibrosis in Cyp2c70-Deficient Mice Are Fully Reversed by Ursodeoxycholic Acid.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJan Freark de Boer,Hilde D de Vries,Anna Palmiotti,Rumei Li,Marwah Doestzada,Joanne A Hoogerland,Jingyuan Fu,Anouk M La Rose,Marit Westerterp,Niels L Mulder,Milaine V Hovingh,Martijn Koehorst,Niels J Kloosterhuis,Justina C Wolters,Vincent W Bloks,Joel T Haas,David Dombrowicz,Bart Staels,Bart van de Sluis,Folkert KuipersPubMed 33309945
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PE/Cy7® Conjugation Kit is a simple and quick process for antibody labeling/conjugation
- Rapid Conjugation: PE/Cy7® labeling in <4 hrs with just 30 secs of hands-on time.
- High Efficiency: Ensures 100% antibody recovery, preserving the integrity and value of your antibodies.
- Versatility: Suitable for conjugating antibodies, proteins, and peptides. The labeled antibodies can be used immediately in applications such as Western Blot, ELISA, and IHC without further purification.
- Confidence: Cited in over 15 publications.
Storage
- Shipped at conditions
- Ambient - Cannot Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
PE/Cy7® Conjugation Kit / PE/Cy7® Labeling Kit ab102903 uses a simple and quick process for PE/Cy7 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our . To conjugate an antibody to PE/Cy7® using this kit: - add modifier to antibody and incubate for 3 hrs - add quencher and incubate for 30 mins The PE/Cy7®conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use. The excitation and emmision wavelengths for PE/Cy7® are Ex: 496nm, Em: 774nm Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid . Use the to learn more about the technology, or about conjugating other proteins and peptides to PE/Cy7®. Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® R-PE/Cy7 Labeling Kit. 762-0005 is the same as the 60 μg size. 762-0010 is the same as the 3 x 60 ug size. 762-0030 is the same as the 3 x 10 ug size. 762-0015 is the same as the 600 μg size.
Amount and volume of antibody for conjugation to PE/Cy7®.
| Kit size | Recommended amount of antibody | Maximum antibody volume1 |
| 3 x 10 μg | 3 x 10 μg | 3 x 10 μL |
| 60 μg | 1 x 60 μg | 1 x 60 μL |
| 3 x 60 μg | 3 x 60 μg | 3 x 60 μL |
| 600 μg | 1 x 600 μg | 1 x 600 μL |
1Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 1 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
The selling size of this product has been changed – it is now based on the amount of antibody that can be conjugated with the kit, not the amount of PE mix provided. The amount of antibody advised that can be used with the kit has also been updated to reflect what will give the best conjugation results. The quantity and formulation of reagents provided have not changed, if you have been previously using the kit successfully with a different amount of antibody, there is no need to change the way that you are using the kit.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Compatible buffer constituents
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.
| 50mM / 0.6% Tris1 | 0.1%/1% BSA2 | 50% glycerol |
| 0.1% sodium azide | PBS | Potassium phosphate |
| Sodium chloride | HEPES | Sucrose |
| Sodium citrate | EDTA | Trehalose |
1 1% BSA gives lower quality conjugates, BSA can also interfere with the use of the conjugated antibody in tissue staining.
Incompatible buffer constituents
| Thiomerosal | Proclin | Glycine |
| Arginine | Glutathione | DTT |
If a constituent of the buffer containing your antibody or protein is not listed above, please contact us.
Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Storing and handling conjugation kits
Lyophilized Lightning-Link® components are hygroscopic.
Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.
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We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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5 product images
Image from Oliveira BM et al., Scientific reports., 9 (1) 3413. Fig 1.; doi: 10.1038/s41598-019-39938-0. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.Conjugation - PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Oliveira BM et al. used ab102903 PE-Cy7® conjugation kit with a mouse anti-bovine CD45RO antibody and APC/Cy7® Conjugation Kit - Lightning-Link® ab102859 APC-Cy7® conjugation kit with a mouse anti-Bovine CD62L antibody. This enabled them to run their desired multicolor flow cytometry panel.
Data shows flow cytometry gating strategy used to define γδ T cells (TCRγδ+CD3+CD335−), CD4+ T cells (CD4+CD3+TCRγδ−CD335−), CD8+ T cells (CD8+CD3+TCRγδ−CD335−) and NK cells (CD335+CD3−) in the stromal vascular fraction (SVF) of mesenteric and subcutaneous bovine adipose tissue (MAT and SAT, respectively) and in peripheral blood leukocytes. Dead cells were excluded with Fixable Viability Dye (FVD), lymphocytes were gated based on SSC-A versus FSC-A and singlets were selected from the FSC-A versus FSC-H dot plot.
The flow cytometry gating strategy used to define CD45RO+ and CD62L+ T cell subpopulations is also shown in CD8+
Image from Flores et al., PloS one, 15(4): e0231977; doi: 10.1371/journal.pone.0231977. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Flores, Erica B et al. used PE/Cy7® Conjugation Kit - Lightning-Link® as part of examining Poxiviruses. They used the kit to conjugate sulfated heparin (clone F58-10E4) for use in flow cytometry.
Sulfation of cell surface heparin measured via flow cytometry utilizing an antibody that recognizes the sulfated 10E4 epitope on HS chains. The mean fluorescent intensity (MFI) values were then normalized to NDST+ values to show the relative change in the NDST-/- cell lines
Image from Robinson, Andrew P., et al., PloS one, 9(9):e107649. doi: 10.1371/journal.pone.0107649. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Flow Cytometry - PE/Cy7 Conjugation Kit- Lightning-Link.
Robinson, Andrew P., et al used PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903) as part of characterizing oligodendroglial populations. They used the kit to conjugate PE/Cy7® to Mouse anti-A2B5 antibody, clone 105, for use in flow cytometry.
SJL/J mice were immunized with PLP139–151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n = 5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45− or CD45low. (B) Oligodendroglial cells were defined by double positive staining: A2B5+PDGFRα+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes.
Image from Okagawa, Tomohiro et al., Vet Res., 49(1):50. doi: 10.1186/s13567-018-0543-9. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Flow Cytometry - PE/Cy7 Conjugation Kit;- Lightning-Link.
Okagawa, Tomohiro, et al used PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903) as part of examining PD-1 and LAG-3 expression. They used the kit to conjugate PE/Cy7® to monoclonal anti-IgM antibody, clone IL-A30, for use in flow cytometry.
Expression of PD-1 and LAG-3 on CD4+ T cells in BLV-infected cattle. A Gating strategy and representative dot plots for expression analyses of PD-1 and LAG-3 on IgM−CD3+CD4+γδTCR− T cells from peripheral blood of BLV-infected cattle (AL and EBL). Values in the quadrants indicate percentages of cells. Percentages of PD-1+LAG-3+CD4+ T cells (B), PD-1+LAG-3−CD4+ T cells (C), and PD-1−LAG-3+CD4+ T cells (D) in CD3+CD4+ T-cell population in peripheral blood from BLV-uninfected (BLV − ; n = 15), AL (n = 22), PL (n = 11), and EBL cattle (n = 7). Bars indicate group median percentage. Significant differences between each group were determined using a Kruskal–Wallis test, where P < 0.05 and P < 0.001, indicated by asterisks (* and ***, respectively).
Image from Nishimori, Asami et al. PloS one vol. 12,4 e0174916. 26 Apr. 2017, doi:10.1371/journal.pone.0174916. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Flow Cytometry - PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Nishimori, Asami et al used PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903) as part of examining bovine leukemia virus infection. They used the kit to conjugate PE/Cy7® to anti-bovine IgM antibody for use in flow cytometry.
A BLV-infected cow (#368, Holstein, female, 538 kg, 31 months old) was inoculated with 530 mg (1 mg/kg) of the purified 4G12 intravenously. (A) The proliferation of CD4+ and CD8+ T cells against BLV antigen. Peripheral blood mononuclear cells (PBMCs) isolated from the cow which was inoculated with 4G12 were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and cultured without stimulation (medium) or with the supernatant of FLK or FLK-BLV cells for 6 days. After the cultivation, the proliferation of T cells was immediately analyzed by flow cytometry. A P-value less than 0.05 was considered statistically significant. #, P <0.05 (FLK-BLV, versus day 0; one-way ANOVA followed by Dunnett's test). (B) Changes in PD-L1 occupancy on circulating IgM+ B cells calculated by the binding of 4G12 to bovine PD-L1. The occupancy was estimated as the percentage of the in vivo PD-L1 binding occurred at the total available binding sites. (C) Changes in BLV provirus loads in the cow inoculated with 4G12; the y-axis shows the number of BLV copies included in 50-ng DNA extracts of PBMCs. Data are means ± SEM of at least three replicate experiments.
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