Product nameAtto 633 Conjugation Kit (Fast) - Lightning-Link®
Atto 633 Conjugation Kit (Fast) - Lightning-Link® (ab269898) provides an easy-to-use, one step procedure that allows researchers to covalently label proteins, peptides and other biomolecules containing primary amines with Atto 633 with only 30 seconds hands-on time; furthermore conjugates are ready to use in less than twenty minutes.
The antibody to be labeled should be purified, in an appropriate buffer for conjugation and at a suitable concentration.
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 Atto 633.
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 Atto 633 Labeling Kit. 353-0015 is the same as the 1 mg size. 353-0010 is the same as the 3 x 100 ug size. 353-0030 is the same as the 3 x 10 ug size. 353-0005 is the same as the 100 µg size.
Amount and volume of antibody for conjugation to Atto 633
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 labeling 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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 µg 1 mg 3 x 10 µg 3 x 100 µg ab274070 - Atto 633 mix (Lyophilized) 1 x 100µg 1 x 1mg 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
Junginger, Johannes et al used Atto 633 Conjugation Kit - Lightning-Link® (ab269898) as part of examining zoonotic intestinal helminths. They used the kit to conjugate Atto 633 to anti-canine CD4 antibody for use in flow cytometry.
Three-colour flow cytometry revealed the TcES-associated increase in Foxp3high lymphocytes to be associated with CD4+, CD4+ CD8+ double-positive and CD4- CD8- double-negative subsets, while this effect was lower in CD8+ T cells. Compared to TcES, treatment with AcES at 150 µg/mL was associated with a much lower elevation in Foxp3high expression by lymphocytes and CD4+ CD8+ T cells.
ab269898 has been referenced in 1 publication.
- Junginger J et al. Zoonotic intestinal helminths interact with the canine immune system by modulating T cell responses and preventing dendritic cell maturation. Sci Rep 7:10310 (2017). PubMed: 28871165