Atto 390 Conjugation Kit (Fast) - Lightning-Link® (ab269895)
Overview
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Product name
Atto 390 Conjugation Kit (Fast) - Lightning-Link® -
Product overview
Atto 390 Conjugation Kit / Atto 390 Labeling Kit ab269895 uses a simple and quick process for Atto 390 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 Atto 390 using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 mins
The Atto 390 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 Atto 390.
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 390 Labeling Kit. 349-0005 is the same as the 100 µg size. 349-0010 is the same as the 3 x 100 ug size. 349-0030 is the same as the 3 x 10 ug size.
Amount and volume of antibody for conjugation to Atto 390
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 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 > 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.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 µg 3 x 10 µg 3 x 100 µg ab274063 - Atto 390 mix (Lyophilized) 1 x 100µg 3 x 10µg 3 x 100µg ab273994 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl ab273995 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl
Images
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Flow Cytometry - Atto 390 Conjugation Kit (Fast) - Lightning-Link (ab269895)Image from Taichman, Russell S., et al., PloS one, 8(4): e61873. doi: 10.1371/journal.pone.0061873. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/
Taichman, Russell S., et al used Atto 390 Conjugation Kit (Fast) - Lightning-Link® (ab269895) as part of examining Axl and Tyro3 expression during experimental prostate cancer (PCa) progression. They used the kit to conjugate Atto 390 to antibodies for use in flow cytometry.
Anti-Axl, anti-Tyro3 and anti-Ki67 antibodies were conjugated to the fluorophores APC-Cy7, PE-Cy5, and Atto390 using our Lightning-Link® Conjugation kits. (A) Experimental model. Human PCa cell lines (PC3Luc, DU145Luc) were implanted s.c. into male SCID mice as a model of a primary (1°) tumor development, and removed after 1 month. At monthly intervals thereafter human PCa cells were identified by anti-HLA staining; proliferative status (Ki67 staining) and Axl or Tyro3 levels were evaluated by FACS. (B) Percent expression of Ki67 by lineage depleted (Lin-) marrow cells or by primary tumor cells at 1 month. (C-D) Percent expression of Axl or Tyro3 by primary tumor cells established with (C) DU145 or (D) PC3 cells or by DTCs recovered from marrow over time.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab269895 has been referenced in 1 publication.
- Taichman RS et al. GAS6 receptor status is associated with dormancy and bone metastatic tumor formation. PLoS One 8:e61873 (2013). PubMed: 23637920