Cy5® conjugation / labeling in < 20 mins with 30 secs hands-on time using Cy5® Conjugation Kit (Fast) - Lightning-Link® ab188288. 100% antibody recovery.
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- Cell metabolism 34:1280-1297.e92022Methionine metabolism controls the B cell EBV epigenome and viral latency.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRui Guo,Jin Hua Liang,Yuchen Zhang,Michael Lutchenkov,Zhixuan Li,Yin Wang,Vicenta Trujillo-Alonso,Rishi Puri,Lisa Giulino-Roth,Benjamin E GewurzPubMed 36070681
- Nature communications 13:43082022Pancreatic tumor eradication via selective Pin1 inhibition in cancer-associated fibroblasts and T lymphocytes engagement.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJiaye Liu,Yang Wang,Chunyang Mu,Meng Li,Kewei Li,Shan Li,Wenshuang Wu,Lingyao Du,Xiaoyun Zhang,Chuan Li,Wei Peng,Junyi Shen,Yang Liu,Dujiang Yang,Kaixiang Zhang,Qingyang Ning,Xiaoying Fu,Yu Zeng,Yinyun Ni,Zongguang Zhou,Yi Liu,Yiguo Hu,Xiaofeng Zheng,Tianfu Wen,Zhihui Li,Yong LiuPubMed 35879297
- Nature communications 13:25402022Spatial epitranscriptomics reveals A-to-I editome specific to cancer stem cell microniches.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmos C Lee,Yongju Lee,Ahyoun Choi,Han-Byoel Lee,Kyoungseob Shin,Hyunho Lee,Ji Young Kim,Han Suk Ryu,Hoe Suk Kim,Seung Yeon Ryu,Sangeun Lee,Jong-Ho Cheun,Duck Kyun Yoo,Sumin Lee,Hansol Choi,Taehoon Ryu,Huiran Yeom,Namphil Kim,Jinsung Noh,Yonghee Lee,Inyoung Kim,Sangwook Bae,Jinhyun Kim,Wooseok Lee,Okju Kim,Yushin Jung,Changhoe Kim,Seo Woo Song,Yeongjae Choi,Junho Chung,Byung Gee Kim,Wonshik Han,Sunghoon KwonPubMed 35534484
- Frontiers in immunology 13:8389662022Discovery of Anti-PD-L1 Human Domain Antibodies for Cancer Immunotherapy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHao Liu,Yanli Liu,Zhen Zhao,Yuanke Li,Bahaa Mustafa,Zhijin Chen,Ashutosh Barve,Akshay Jain,Xiaolan Yao,Guangfu Li,Kun ChengPubMed 35444660
- Cell reports 38:1104112022Epstein-Barr virus BNRF1 destabilizes SMC5/6 cohesin complexes to evade its restriction of replication compartments.Applications:Unspecified applicationReactive species:Unspecified reactive speciesStephanie Pei Tung Yiu,Rui Guo,Cassie Zerbe,Michael P Weekes,Benjamin E GewurzPubMed 35263599
- Frontiers in immunology 12:7137042021IgG Immune Complexes Inhibit Naïve T Cell Proliferation and Suppress Effector Function in Cytotoxic T Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWissam Charab,Matthew G Rosenberger,Haridha Shivram,Justin M Mirazee,Moses Donkor,Soumya R Shekhar,Donjeta Gjuka,Kimberly H Khoo,Jin Eyun Kim,Vishwanath R Iyer,George GeorgiouPubMed 34447380
- Frontiers in bioengineering and biotechnology 8:6066522021Characterization of the Biodistribution of a Silica Vesicle Nanovaccine Carrying a Protective Antigen With Live Animal Imaging.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKarishma T Mody,Bing Zhang,Xun Li,Nicholas L Fletcher,Dewan T Akhter,Sandy Jarrett,Jun Zhang,Chengzhong Yu,Kristofer J Thurecht,Timothy J Mahony,Neena MitterPubMed 33537291
- Cell 184:1000-1016.e272021Functional characterization of the dural sinuses as a neuroimmune interface.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJustin Rustenhoven,Antoine Drieu,Tornike Mamuladze,Kalil Alves de Lima,Taitea Dykstra,Morgan Wall,Zachary Papadopoulos,Mitsuhiro Kanamori,Andrea Francesca Salvador,Wendy Baker,Mackenzie Lemieux,Sandro Da Mesquita,Andrea Cugurra,James Fitzpatrick,Sanja Sviben,Ross Kossina,Peter Bayguinov,Reid R Townsend,Qiang Zhang,Petra Erdmann-Gilmore,Igor Smirnov,Maria-Beatriz Lopes,Jasmin Herz,Jonathan KipnisPubMed 33508229
- Materials (Basel, Switzerland) 13:2020Tubular Dentin Regeneration Using a CPNE7-Derived Functional Peptide.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYoon Seon Lee,Yeoung-Hyun Park,Dong-Seol Lee,You-Mi Seo,Ji-Hyun Lee,Joo-Hwang Park,Han-Wool Choung,So-Hyun Park,Won Jun Shon,Joo-Cheol ParkPubMed 33081300
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Cy5® conjugation / labeling in < 20 mins with 30 secs hands-on time using Cy5® Conjugation Kit (Fast) - Lightning-Link® ab188288. 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
Cy5® Conjugation Kit / Cy5® Labeling Kit (ab188288) uses a simple and quick process for Cy5 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our . To conjugate an antibody to Cy5 using this kit: - add modifier to antibody and incubate for 15 min - add quencher and incubate for 5 mins The Cy5 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 Cy5. 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® Rapid Cy5 Labeling Kit. 342-0005 is the same as the 100 μg size. 342-0010 is the same as the 3 x 100 ug size. 342-0030 is the same as the 3 x 10 ug size. 342-0015 is the same as the 1 mg size.
Amount and volume of antibody for conjugation to Cy5
| Kit size | Recommended amount of antibody1 | Maximum amount of antibody | Maximum antibody volume2 |
| 3 x 10 μg | 3 x 10 μg | 3 x 20 μg | 3 x 10 μL |
| 100 μg | 1 x 100 μg | 1 x 200 μg | 1 x 100 μL |
| 3 x 100 μg | 3 x 100 μg | 3 x 200 μg | 3 x 100 μL |
| 1 mg | 1 x 1 mg | 1 x 2 mg | 1 x 1 mL |
1 Using the maximum amount of antibody may result in less labelling per antibody.
2 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 2mg/ml or < 0.5 mg/ml should be diluted /concentrated.
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% BSA2 | 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%
2 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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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:
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6 product images
Image from Lee et al., Materials, 13(20):4618; doi: 10.3390/ma13204618. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Immunocytochemistry - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Lee, Yoon Seon, et al used Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) as part of examining the translocation and localization of newly developed peptide derived from CPNE7 (Cpne7-DP) . They used the kit to conjugate Cy5® to Cpne7-DP oligopeptide for use in immunocytochemistry.
The intracellular distribution of Cpne7-DP is shown after odontoblastic MDPC-23 cells were treated with Cy5-labeled Cpne7-DP (10 μg/mL).
Image from Lindenburg, Laurens, et al., Nucleic acids res., 8(11): e63; doi: 10.1093/nar/gkaa270. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Flow Cytometry - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Lindenburg, Laurens, et al used Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) as part of examining microemulsion-based bead display screening of the ZIgE SpliMLiB library. They used the kit to conjugate Cy5® to Native human IgE protein (Azide free) (Human IgE protein (Azide free) ab65866) for use in FACS.
(A) Schematic overview of a round of SpliMLiB-enabled directed evolution of ZIgE. SpliMLiB beads (i) were singly encapsulated in emulsion IVTT at 37°C for 1 h (ii), sufficient time to allow for both ZIgE-SpyCatcher variants' expression as well as for their SpyTag-SpyCatcher-mediated immobilisation on the bead surface, after which the emulsion was broken, and the washed beads were exposed to Cy5-labelled IgE (iii), followed by flow cytometric sorting of beads based on Cy5 signal (iv). (B) Representative histogram recorded during the flow cytometric sorting of SpliMLiB ZIgE library beads. The range of fluorescence intensity used for each of the sorting gates 1-4 is indicated. (C) Analysis of pooled, recovered and subcloned DNA from the sorting gates set out in panel B. DNA was used to express protein in IVTT under bulk, i.e. non-emulsion conditions, in the presence of SpyTag-functionalised microbeads. The microbeads, having captured the SpyCatcher fusion proteins, were then incubated with 200 nM IgE-Cy5 and analysed by flow cytometry. Cy5 fluorescence intensity was normalised to a sample prepared from beads exposed to purified ZIgEwild-type-SpyCatcher protein (WT, grey bar). Negative control (NC) was beads not exposed to any ZIgE-SpyCatcher protein. (D) Analysis of bacterially expressed & purified variants derived from the stringently sorted library output from FACS sorting gate 4. Beads that had been bound with ZIgE-SpyCatcher variants were incubated with 200 nM IgE-Cy5 and analysed by flow cytometry. ZIgEwild-type-SpyCatcher (labelled WT) was included as control and was used to normalise all fluorescent values. The variant showing the highest Cy5 median signal (variant 33, marked by a single asterisk) and second highest (variant 44, marked by a double asterisk) signal were taken forward for further analysis. (E) As panel D, except for 48 randomly picked clones derived from the unsorted SpliMLiB input library beads. (F) Frequencies of amino acids encountered in selected variants displaying a higher binding signal than ZIgEwild-type-SpyCatcher (17 in total). The most frequent amino acid at each position is indicated in bold to emphasise it.
Image from Mody et al., Front bioeng biotechnol., 8:606652; doi: 10.3389/fbioe.2020.606652. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Western blot - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Mody, Karishma T., et al used Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) as part of examining the integrity of Bm86 protein postlabeling. They used the kit to conjugate Cy5® to R. micr-protein ladder, lane 5-Cy5-Bm86 protein, lane 7-Cy5-Bm86/ Rho-SV-140-C18 Supernatant, lane 8-Cy5-Bm86/Rho-SV-140-C18 pellet (first antibody 869 polyclonal sheep 1/5,000; second antibody monoclonal anti-goat/sheep 1/10,000). The Cy5-labeled Bm86 retained its native antigenicity as it was recognized by the antibodies in serum from a sheep immunized with the unlabeled antigen from a previous study.
Image from Mellal et al., Sci rep., 9(1):12903. doi: 10.1038/s41598-019-49472-8. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/FRET - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Mellal, Katia, et al used Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) as part of examining effects of MPE-001 on the CD36/TLR2 interaction. They used the kit to conjugate Cy5® to anti-CD36 antibody (Anti-CD36 antibody [MF3] - Low endotoxin, Azide free ab80080) for use in FRET.
Peritoneal MPs were stimulated with 300 ng/ml R-FSL1 in the presence of 10-7 M MPE-001 or vehicle. (A) MPE-001 disrupted the interaction between CD36 labeled with Cy5 (red) using the Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) and TLR2 labeled with Cy3 (green) using the Cy3® Conjugation Kit (Fast) - Lightning-Link® (Cy3® Conjugation Kit (Fast) - Lightning-Link® ab188287) as assessed by FRET after 5 min stimulation with R-FSL1. (B) Percentage of energy transfer measured using LSM-700 confocal microscope (Zeiss). One-way ANOVA test with Newman-Keuls post-test for multiple comparison was performed. *P<0.05, **P
Image from Li, Tiankuan, et al., Biomaterials science, 8(5):1418-1430. doi: 10.1039/c9bm01575b. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/3.0/Fluorescence Microscopy - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Li, Tiankuan, et al used Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288) as part of examining immunotherapeutical treatments for lung cancer. They used the kit to conjugate Cy5® to anti-PD-L1 mAb for use in fluorescence microscopy.
CLSM images of docetaxel and CY5-labelled anti-PD-L1 mAb-co-loaded microbubbles with a shell containing FITC, scale bar = 20 μm.
Image from Dassanayake RP et al., PloS One, 12(8):e0183610. Fig 4.; doi:10.1371/journal.pone.0183610. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Immunocytochemistry/ Immunofluorescence - Cy5® Conjugation Kit (Fast) - Lightning-Link® (ab188288)
Cy5® Conjugation Kit - Lightning-Link® labeling NK2A peptide for confocal microscopy.
Dassanayake, Rohana P et al used ab188288 as part of examining the antimicrobial activity of bovine NK-lysin-derived peptides.
They used the kit to conjugate Cy5® to NK2A peptide for use in confocal microscopy.
H. somni was incubated with unconjugated Cy5 (A) or 20 μM NK2A-Cy5 conjugate for 30 min and bacterial nuclear contents were stained with DAPI. NK2A-Cy5 conjugate is shown in red and DAPI is shown in blue.
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