Product nameDyLight® 594 Conjugation Kit (Fast) - Lightning-Link®
DyLight® 594 Conjugation Kit / DyLight 594 Labeling Kit (ab201801) uses a simple and quick process for DyLight 594 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.
To conjugate an antibody to DyLight® 594 using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 mins
The DyLight® 594 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® 594.
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® 594 Labeling Kit. 324-0015 is the same as the 1 mg size. 324-0030 is the same as the 3 x 10 ug size. 324-0010 is the same as the 3 x 100 ug size. 324-0005 is the same as the 100 ug size.
Amount and volume of antibody for conjugation to Dylight® 594
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 ab274019 - Dylight® 594 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
Hsiao, Feng, et al used DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (ab201801) as part of examining SAMHD1 expression in tissue Tm subsets. They used the kit to conjugate DyLight® 594 to anti-SAMHD1 antibody for use in flow cytometry.
A) Validation of SAMHD1 antibody by flow cytometry. Unstimulated PBMCs were mock-treated or exposed for 3 days to F4.HSA. Depicted within the gates are the proportions of infected cells amongst the SAMHD1- and SAMHD1+ cells. The results demonstrate that SAMHD1- cells, which are permissive, harbor a higher proportion of infected cells than non-permissive, SAMHD1+ cells, as expected . Results are pre-gated on live, singlet CD3+CD8-CD45RO+CD45RA- cells. B) Bar graph of the percentage of SAMHD1+ cells across the three indicated memory CD4+ T cell subsets from HLACs as determined by flow cytometry. Results are pre-gated on live, singlet CD3+CD8-CD45RO+CD45RA- cells. ns: non-significant as determined using a 2-tailed paired parametric t-test.
Lalo, Ulyana, et al used DyLight® 594 Conjugation Kit (Fast) - Lightning-Link® (ab201801) as part of examining the colocalization of vesicular ATP transporters with exocytotic organelle markers. They used the kit to conjugate DyLight® 594 to various antibodies for use in multiphoton fluorescent microscopy.
Living, acutely isolated cortical astrocytes were labeled with antibodies to vesicular transporters (VNUT1 and VGLUT1) and synaptic vesicle (SV2A) and lysosomal (LAMP3 and Cathepsin-D) markers. Antibodies were conjugated to fluorescent dyes DyLight488 (green) and DyLight594 (red) prior to astrocyte labeling. (A-D) The representative two-photon fluorescence images (maximal intensity projections of Z-stack) and results of colocalization analysis, carried out using NIH ImageJ 1.43 software. The correlation between green and red fluorescence (images in the right column) is depicted as a product of the relative differences from the mean (PDM) for each pixel; the pseudocolor PDM images were generated as an output of ImageJ analysis routine. Positive values (bright yellow) are indicative for good co-localization of green and red signals, negative values (blue-violet) indicate segregation, and black color shows the lack of correlation. Note the different extent of scale for PDM values in (A-D). All scale bars in (A-D) are 5 µm.
ab201801 has been referenced in 5 publications.
- Vessey KA et al. Fluorescent Labeling and Quantification of Vesicular ATP Release Using Live Cell Imaging. Methods Mol Biol 2041:209-221 (2020). PubMed: 31646491
- Hsiao F et al. Tissue memory CD4+ T cells expressing IL-7 receptor-alpha (CD127) preferentially support latent HIV-1 infection. PLoS Pathog 16:e1008450 (2020). PubMed: 32353080
- Kaushik DK et al. Enhanced glycolytic metabolism supports transmigration of brain-infiltrating macrophages in multiple sclerosis. J Clin Invest 129:3277-3292 (2019). PubMed: 31112527
- Adam A et al. Multiplexed FluoroSpot for the Analysis of Dengue Virus- and Zika Virus-Specific and Cross-Reactive Memory B Cells. J Immunol 201:3804-3814 (2018). PubMed: 30413671
- Lalo U et al. Exocytosis of ATP from astrocytes modulates phasic and tonic inhibition in the neocortex. PLoS Biol 12:e1001747 (2014). PubMed: 24409095