PE/Cy5.5® Conjugation Kit - Lightning-Link®
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(10 Publications)
- Rapid Conjugation: achieve PE/Cy5.5 labeling in under 4 hours with just 30 seconds of hands-on time.
- High Efficiency: ensures 100% antibody recovery, meaning no loss of valuable antibodies.
- Versatility: suitable for conjugating antibodies, proteins, and peptides. PE/Cy5.5 labeled antibodies can be used immediately in applications such asFlow cytometry, ELISA, and Immunohistochemistry (IHC) without further purification.
- Flow Cyt
PubMed
Flow Cytometry - PE/Cy5.5® Conjugation Kit - Lightning-Link® (AB102899)
Sancey, Lucie, et al used PE/Cy5.5® Conjugation Kit - Lightning-Link® (ab102899) as part of examining AGuIX NP uptake measured by FCM as a function of time in the 3 OncoCilAir™ subpopulations. They used the kit to conjugate PE/Cy5.5® to IgG isotype control antibody for use in flow cytometry.
a Summary of AGuIX® uptake with the percentage of AGuIX®+ cells and their corresponding MFI values presented after 24 and 72 h of NP exposure (n=6/conditions, from 3 distinct batches of cell culture with and without mucus). The relative uptake ratios were calculated from Eq. 3. Statistical analysis was performed using the Mann-Whitney test (*p 0.05 compared to GFP-negative cells). The results b-i were obtained from samples of the 2nd series of cell cultures, presenting a fast tumor growth and an intense AGuIX® uptake at T24h. FACS plots of AGuIX® uptake (% of AGuIX+-cells and geometric mean MFI) according to the 3 gates (b, f) that discriminate GFP-negative (red channel; c, g), GFP+ (green channel; d, h) and GFP++ (blue channel; e, i) tumor cells at 24 h (b-e) and 72 h (f-i)
Image from Sancey, Lucie, et al., J. nanobiotechnology, 18(1):129; doi: 10.1186/s12951-020-00683-6. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
- Schematic Diagram
Supplier Data
Schematic Diagram - PE/Cy5.5® Conjugation Kit - Lightning-Link® (AB102899)
This illustration demonstrates a general procedure of how Lightning-Link® labeling technology enables the direct labeling of antibodies or proteins.
Simply pipette your antibody or biomolecule of choice into the vial of a lyophilized mixture containing the label of interest and incubate for around 3 hours. Please see the ab102899 protocol booklet for more details.
Learn more about our Lightning-Link® conjugation kits here
- Flow Cyt
PubMed
Flow Cytometry - PE/Cy5.5® Conjugation Kit - Lightning-Link® (AB102899)
Wang, Shuyan, et al used PE/Cy5.5® Conjugation Kit - Lightning-Link® (ab102899) as part of examining Neph3-positive cells co-cultured with rat cerebellar slices. They used the kit to conjugate PE/Cy5.5® to anti-Neph3 monoclonal antibody for use in flow cytometry.
(A) Schematic representation of sorting and co-culture procedures. (B, C) FACS analysis of neural rosette cells on Day 20 of differentiation. (D) Immunofluorescence staining to confirm that the sorted cells were Neph3/GFP-double positive. Bar = 100 μm. (E) The sorted cells were co-cultured with cerebellum slices and examined at 1 week and 4 weeks. Bar = 100 μm.
Image from Wang, Shuyan, et al., Scientific reports, 5:9232; doi: 10.1038/srep09232. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Product details
PE/Cy5.5® Conjugation Kit - Lightning-Link® (ab102899) uses a simple and quick process for PE/Cy5.5 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides.
To conjugate an antibody to PE/Cy5.5® using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The PE/Cy5.5® conjugated antibody can be used immediately in Flow cytometry and other applications. No further purification is required and 100% of the antibody is recovered for use.
Learn about our antibody labeling kits and their advantages here.
Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our antibody purification and concentration kits.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
Amount and volume of antibody for conjugation to PE/Cy5.5®.
| 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% BSA | 50% glycerol |
| 0.1% sodium azide | PBS | Potassium phosphate |
| Sodium chloride | HEPES | Sucrose |
| Sodium citrate | EDTA | Trehalose |
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
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.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® R-PE/Cy5.5 Labeling Kit. 761-0005 is the same as the 60 µg size. 761-0010 is the same as the 3 x 60 µg size. 761-0030 is the same as the 3 x 10 µg size. 761-0015 is the same as the 600 µg size.
What's included?
Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
- Download websiteProtocolBooklet|en
- Download websiteProtocolBooklet|zh
Target data
Publications (10)
Recent publications for all applications. Explore the full list and refine your search
Current protocols 2:e589 PubMed36373983
2022
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Cell reports 34:108863 PubMed33691089
2021
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Med (New York, N.Y.) 2:243-262.e8 PubMed33521747
2021
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medRxiv : the preprint server for health sciences : PubMed32935120
2020
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Journal of nanobiotechnology 18:129 PubMed32912214
2020
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Cell 182:625-640.e24 PubMed32702313
2020
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Genetics in medicine : official journal of the Ame 19:1071-1077 PubMed28301456
2017
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Scientific reports 5:9232 PubMed25782665
2015
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FASEB journal : official publication of the Federation of American Societies for Experimental Biology 26:1301-10 PubMed22121051
2011
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Nature medicine 16:446-51 PubMed20208541
2010
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com