Product nameDyLight® 550 Conjugation Kit (Fast) - Lightning-Link®
DyLight® 550 Fast Conjugation Kit (ab201800) uses a simple and quick process to conjugate an antibody to DyLight® 550. It can also be used to conjugate other proteins or peptides.
This product is manufactured by Expedeon, an Abcam company. See the table below to match Abcam kit size against Expedeon product code.
To conjugate an antibody to DyLight® 550 using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 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 antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to DyLight® 550.
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 DyLight® 550 Labeling Kit. 323-0015 is the same as the 1 mg size. 323-0010 is the same as the 3 x 100 ug size. 323-0030 is the same as the 3 x 10 ug size. 323-0005 is the same as the 100 ug size.
Amount and volume of antibody for conjugation to DyLight® 550
Kit size Recommended
amount of antibody1
amount of antibody
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
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.
DyLight® is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg ab274016 - Dylight® 550 Conjugation Mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg ab273994 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab273995 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
Soldevila, Ferran, et al used DyLight® 550 Conjugation Kit (Fast) - Lightning-Link® (ab201800) as part of examining myeloid cell populations in the porcine palatine tonsil. They used the kit to conjugate DyLight® 550 to anti-CD14 IgG for use in confocal microscopy.
In situ localization of the CD14+ cells and plasmocytoid dendritic cells in porcine palatine tonsil. CD14+ cells and pDCs were localized by confocal microscopy following ethanol fixation of tonsil slices. The areas assessed included the follicle (F), the interfollicular region (IFA), the crypt (C). The tissue was stained using panel 1 antibodies; white arrow CD14+ cells and yellow arrow pDCs. Images are representative of at least two images from each section, from three different pigs. Objective used: (A) 63× oil immersion. Scale bars as shown.
Lucas, Nicolas, et al used DyLight® 550 Conjugation Kit (Fast) - Lightning-Link® (ab201800) as part of examining the function of α-melanocyte-stimulating hormone (α-MSH) in Melanocortin 4 receptor (MC4R) signaling in normal and pathological conditions. They used the kit to conjugate DyLight® 550 to anti-α-MSH IgG for use in confocal microscopy.
Representative images of hMC4R GFP+ HEK 293 cells (green) 30 min after application of DyLight 550®-labeled (red) α-MSH affinity-purified IgG from eating disorder (anorexia nervosa, n = 9; bulimia nervosa, n = 7; binge eating disorder, n = 7), obese (n = 10), and Ctrl (n = 9) preincubated or not with α-MSH.
Adult mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS.
Sections were then incubated with either:
(A) Mouse Anti-DDX4 (ab27591) at a 1:500 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Donkey Anti-Mouse 555 (ab150110) applied at a 1:500 dilution.
(B) Mouse Anti-DDX4 (ab27591) labeled with Dylight 550 Fast Conjugation Kit (ab201800). Sections were incubated with labeled antibody at 1:100 dilution for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and mounted.
DDX4 is shown in red. Phalloidin, which stains F-actin is shown in blue.
ab201800 has been referenced in 9 publications.
- Shin W et al. Dysfunction of Hair Follicle Mesenchymal Progenitors Contributes to Age-Associated Hair Loss. Dev Cell 53:185-198.e7 (2020). PubMed: 32315612
- Lucas N et al. Immunoglobulin G modulation of the melanocortin 4 receptor signaling in obesity and eating disorders. Transl Psychiatry 9:87 (2019). PubMed: 30755592
- Bar-Yoseph H et al. Infliximab-Tumor Necrosis Factor Complexes Elicit Formation of Anti-Drug Antibodies. Gastroenterology 157:1338-1351.e8 (2019). PubMed: 31401142
- Mihklepp K et al. Immunodetection of Streptococcus uberis pathogen in raw milk. Enzyme Microb Technol 130:109360 (2019). PubMed: 31421723
- Niedenberger BA & Geyer CB Advanced immunostaining approaches to study early male germ cell development. Stem Cell Res 27:162-168 (2018). PubMed: 29475796
- Chen CY et al. Suppression of detyrosinated microtubules improves cardiomyocyte function in human heart failure. Nat Med 24:1225-1233 (2018). PubMed: 29892068
- Soldevila F et al. Characterization of the Myeloid Cell Populations' Resident in the Porcine Palatine Tonsil. Front Immunol 9:1800 (2018). PubMed: 30158925
- Mihklepp K et al. Design and production of antibodies for the detection of Streptococcus uberis. Enzyme Microb Technol 96:135-142 (2017). PubMed: 27871373
- Jiang P et al. Histones facilitate a-synuclein aggregation during neuronal apoptosis. Acta Neuropathol 133:547-558 (2017). PubMed: 28004278