DyLight® 650 Conjugation Kit (Fast) - Lightning-Link® (ab201803)
Overview
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Product name
DyLight® 650 Conjugation Kit (Fast) - Lightning-Link® -
Product overview
DyLight® 650 Conjugation Kit / DyLight® 650 Labeling Kit ab201803 uses a simple and quick process for DyLight 650 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.
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® 650 using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 mins
The DyLight 650 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® 650.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
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Notes
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid DyLight® 650 Labeling Kit. 326-0015 is the same as the 1 mg size. 326-0010 is the same as the 3 x 100 ug size. 326-0030 is the same as the 3 x 10 ug size. 326-0005 is the same as the 100 ug size.
Amount and volume of antibody for conjugation to Dylight® 650
Kit size Recommended
amount of antibody1Maximum
amount of antibodyMaximum antibody
volume23 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 > 2 mg/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 check the FAQ or 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.
DyLight® is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg ab274025 - Dylight® 650 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 -
Research areas
Associated products
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Related Products
Images
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Flow Cytometry - DyLight<;650 Conjugation Kit (Fast)- Lightning-Link (ab201803)Image from Saha, Chaitrali, et al., Front Immunol., 8:275. doi: 10.3389/fimmu.2017.00275. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Saha, Chaitrali, et al used DyLight® 650 Conjugation Kit (Fast) - Lightning-Link® (ab201803) as part of examining the ability of monomeric IgA (mIgA) isolated from pooled plasma of healthy donors to modulate human Th17 cells. They used the kit to conjugate DyLight® 650 to IgA and IgG for use in flow cytometry.
Monomeric IgA (mIgA) binds to CD4+ T cells. (A,B) Representative dot plots and percentage (mean ± SEM, n = 3–9 donors) of binding of DyLight650-conjugated mIgA to CD4+ T cells. Statistical significance as determined by two-tailed Student’s t-test is indicated (**P < 0.01). (C,D) Representative dot plots and percentage (mean ± SEM, n = 4) of binding of DyLight650-conjugated mIgA to CD4+ T cells, CD4+CD45RA+ naïve T cells, and CD4+CD45RO+ memory T cells. Statistical significance as determined by one-way ANOVA is indicated (**P < 0.01; ***P < 0.001; ns, not significant). -
Fluorescence Microscopy - DyLight Lightning-LinkImage from Jiang, Peizhou, et al., Scientific reports 7.1 (2017): 1-13. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/Jiang, Peizhou, et al used DyLight® 650 Conjugation Kit - Lightning-Link® (ab201803) as part of examining α-synuclein aggregates. They used the kit to conjugate DyLight® 650 to α-synuclein protein aggregates for use in confocal microscopy.
Membrane penetration- and endocytosis-mediated αS seeding contribute to formation of endo-lysosome-free and endo-lysosome-associated αS inclusions. (a) H4/V1S-SV2/LAMP1-eCFP cells were respectively treated with DyLight® 650 labeled αS seeds (Agg. αS), Dynasore, and Dynasore & Agg. αS for 2 days followed by imaging under confocal microscopy. The concentration for Agg. αS is 0.5 µg/ ml, and Dynasore 20 µM. Yellow and white arrows denote αS seeds and induced LAMP1-free (yellow), LAMP1-positive (white) αS inclusion, respectively. Scale bar: 5 µm. (b) Bar graphs show the comparisons among Agg. αS, Dynasore, and Dynasore & Agg. αS treated cells in the ratio of cells with inclusions to total cells, inclusions to total seeds, LAMP1-positive seeds to total seeds, LAMP1-negative and LAMP1-positive inclusions to total inclusions, respectively. Error bars represent standard error of the mean (N.S = non-significant; **p < 0.01, comparing subsets linked by line, n = 3). -
Conjugation - DyLight® 650 Fast Conjugation Kit (ab201803)This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (12)
ab201803 has been referenced in 12 publications.
- Jiang P et al. Apoptotic Neuron-Derived Histone Amyloid Fibrils Induce a-Synuclein Aggregation. Mol Neurobiol 58:867-876 (2021). PubMed: 33048264
- Galeotti C et al. Intravenous immunoglobulin induces IL-4 in human basophils by signaling through surface-bound IgE. J Allergy Clin Immunol 144:524-535.e8 (2019). PubMed: 30529242
- Swärd K et al. Identification of the intermediate filament protein synemin/SYNM as a target of myocardin family coactivators. Am J Physiol Cell Physiol 317:C1128-C1142 (2019). PubMed: 31461342
- Learn GD et al. Woven collagen biotextiles enable mechanically functional rotator cuff tendon regeneration during repair of segmental tendon defects in vivo. J Biomed Mater Res B Appl Biomater N/A:N/A (2018). PubMed: 30485649
- 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