Labeling in < 4 hrs with 30 secs hands-on time using PE/Cy5.5® Conjugation Kit - Lightning-Link® ab102899. 100% antibody recovery.
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- Current protocols 2:e5892022Do more with Less: Improving High Parameter Cytometry Through Overnight Staining.Applications:Unspecified applicationReactive species:Unspecified reactive speciesCarly E Whyte,Damon J Tumes,Adrian Liston,Oliver T BurtonPubMed 36373983
- Cell reports 34:1088632021Metabolic programs define dysfunctional immune responses in severe COVID-19 patients.Applications:Unspecified applicationReactive species:Unspecified reactive speciesElizabeth A Thompson,Katherine Cascino,Alvaro A Ordonez,Weiqiang Zhou,Ajay Vaghasia,Anne Hamacher-Brady,Nathan R Brady,Im-Hong Sun,Rulin Wang,Avi Z Rosenberg,Michael Delannoy,Richard Rothman,Katherine Fenstermacher,Lauren Sauer,Kathyrn Shaw-Saliba,Evan M Bloch,Andrew D Redd,Aaron A R Tobian,Maureen Horton,Kellie Smith,Andrew Pekosz,Franco R D'Alessio,Srinivasan Yegnasubramanian,Hongkai Ji,Andrea L Cox,Jonathan D PowellPubMed 33691089
- Med (New York, N.Y.) 2:243-262.e82021A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlyssa Silva-Cayetano,William S Foster,Silvia Innocentin,Sandra Belij-Rammerstorfer,Alexandra J Spencer,Oliver T Burton,Sigrid Fra-Bidó,Jia Le Lee,Nazia Thakur,Carina Conceicao,Daniel Wright,Jordan Barrett,Nicola Evans-Bailey,Carly Noble,Dalan Bailey,Adrian Liston,Sarah C Gilbert,Teresa Lambe,Michelle A LintermanPubMed 33521747
- medRxiv : the preprint server for health sciences :2020Metabolic programs define dysfunctional immune responses in severe COVID-19 patients.Applications:Unspecified applicationReactive species:Unspecified reactive speciesElizabeth A Thompson,Katherine Cascino,Alvaro A Ordonez,Weiqiang Zhou,Ajay Vaghasia,Anne Hamacher-Brady,Nathan R Brady,Im-Hong Sun,Rulin Wang,Avi Z Rosenberg,Michael Delannoy,Richard Rothman,Katherine Fenstermacher,Lauren Sauer,Kathyrn Shaw-Saliba,Evan M Bloch,Andrew D Redd,Aaron Ar Tobian,Maureen Horton,Kellie Smith,Andrew Pekosz,Franco R D'Alessio,Srinivasan Yegnasubramanian,Hongkai Ji,Andrea L Cox,Jonathan D PowellPubMed 32935120
- Journal of nanobiotechnology 18:1292020Multiparametric investigation of non functionalized-AGuIX nanoparticles in 3D human airway epithelium models demonstrates preferential targeting of tumor cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLucie Sancey et. al.PubMed 32912214
- Cell 182:625-640.e242020Microglia Require CD4 T Cells to Complete the Fetal-to-Adult Transition.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEmanuela Pasciuto et. al.PubMed 32702313
- Genetics in medicine : official journal of the Ame 19:1071-10772017Prediction of breast cancer risk based on flow-variant analysis of circulating peripheral blood B cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMahrukh M Syeda et. al.PubMed 28301456
- Scientific reports 5:92322015Differentiation of human induced pluripotent stem cells to mature functional Purkinje neurons.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShuyan Wang et. al.PubMed 25782665
- FASEB journal : official publication of the Federation of American Societies for Experimental Biology 26:1301-102011Distinctive immunoregulatory effects of adenosine on T cells of older humans.Applications:Unspecified applicationReactive species:Unspecified reactive speciesCharles S Hesdorffer,Enkhzol Malchinkhuu,Arya Biragyn,Omar S Mabrouk,Robert T Kennedy,Karen Madara,Dennis D Taub,Dan L Longo,Janice B Schwartz,Luigi Ferrucci,Edward J GoetzlPubMed 22121051
- Nature medicine 16:446-512010HIV-1 infects multipotent progenitor cells causing cell death and establishing latent cellular reservoirs.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChristoph C Carter et. al.PubMed 20208541
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Labeling in < 4 hrs with 30 secs hands-on time using PE/Cy5.5® Conjugation Kit - Lightning-Link® ab102899. 100% antibody recovery.
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/Cy5.5® Conjugation Kit / PE/CY5.5® Labeling Kit B-Phycoerythrin Conjugation Kit - Lightning-Link® ab102871 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. Learn about our . 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 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. 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/Cy5.5®. 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/Cy5.5 Labeling Kit. 761-0005 is the same as the 60 μg size. 761-0010 is the same as the 3 x 60 ug size. 761-0030 is the same as the 3 x 10 ug size. 761-0015 is the same as the 600 μg size.
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.
Product promise
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.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
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/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 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/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.
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