Atto 633 Conjugation Kit (Fast) - Lightning-Link® (ab269898)
Overview
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Product name
Atto 633 Conjugation Kit (Fast) - Lightning-Link® -
Product overview
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.
The excitation and emmision wavelengths for Atto 633 are Ex: 630nm, Em: 651nm.
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.
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Notes
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 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 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 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.
Properties
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Storage instructions
Store 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
Images
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Flow Cytometry - Atto 633 Conjugation Kit (Fast) - Lightning-Link (ab269898)Image from Junginger, Johannes et al., Scientific reports vol. 7,1 10310. 4 Sep. 2017, doi:10.1038/s41598-017-10677-4. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
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. -
Atto 633 Conjugation Kit (Fast) - Lightning-Link® absorbance emission graph
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab269898 has been referenced in 7 publications.
- Dou C et al. Sialylation of TLR2 initiates osteoclast fusion. Bone Res 10:24 (2022). PubMed: 35232979
- Chen CY et al. Suppression of detyrosinated microtubules improves cardiomyocyte function in human heart failure. Nat Med 24:1225-1233 (2018). PubMed: 29892068
- 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
- Chang YF et al. The utility of a high-throughput scanning biosensor in the detection of the pancreatic cancer marker ULBP2. Biosens Bioelectron 41:232-7 (2013). PubMed: 22959016
- Su LC et al. Binding kinetics of biomolecule interaction at ultralow concentrations based on gold nanoparticle enhancement. Anal Chem 83:3290-6 (2011). PubMed: 21466206
- Chang YF et al. Discrimination of breast cancer by measuring prostate-specific antigen levels in women's serum. Anal Chem 83:5324-8 (2011). PubMed: 21591802
- Wauters J et al. Flow cytometric detection of myeloperoxidase in horse neutrophils: a novel technique in equine diagnostic research. Vet Immunol Immunopathol 144:417-22 (2011). PubMed: 22018886