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Invitrogen™ SiteClick™ sDIBO Alkyne Kits for Antibody Labeling
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Description
Generate site-specific labeled antibody conjugates with SiteClick sDIBO Alkynes for Antibody Labeling. Our SiteClick sDIBO Antibody Labeling kits are optimized to be used in conjunction with the SiteClick Azido Modification kits (S20026, S10900, or S10901 sold separately) and contain sufficient reagents to perform a conjugation reaction of a SiteClick sDIBO Alkyne label using 100 μg, 1 mg, or 5 mg of azido-modified IgG antibody. The SiteClick sDIBO Alkynes for Antibody Labeling employs copper-free click chemistry to covalently link a dibenzocyclooctyne (sDIBO) labeled Alexa Fluor dye, pHrodo dye, or biotin to an azido modified antibody. The label, when attached to the antibody, is located far away from its antigen binding domain.
Create your own covalently labeled antibody conjugates using the SiteClick sDIBO Alkyne labels, which are intended for use with azido-modified antibodies. SiteClick sDIBO Alkyne Antibody Labeling Kits include the sDIBO Alkyne and the antibody concentrator, enabling copper-free covalent attachment of Alexa Fluor and pHrodo dyes, biotin, or molecules of your choice. When used with the SiteClick Antibody Azido Modification Kit (available separately as Cat. No. S20026, S10900, S10901), labeling occurs on the heavy chain N-linked glycans, far away from the antigen-binding domain, enabling excellent reproducibility between separate labeling reactions and multiple antibody species and subtypes. The SiteClick sDIBO alkyne kits for antibody labeling are optimized for conjugation reactions, covalently linking 100 μg, 1 mg, or 5 mg of azide-modified IgG antibody.
The SiteClick Antibody Azido Modification Kit (Cat. No. S20026, S10900, S10901) is recommended for the attachment of an azide moiety to the heavy chains of an unlabeled IgG antibody. Other engineered antibodies containing azido moieties are also suitable.
When used in conjunction with the SiteClick Antibody Azido Modification Kit, the SiteClick sDIBO alkyne kits for antibody labeling give you the option of trying different fluorescent labels for your antibody, attaching other molecules via streptavidin, or attaching your own molecule via amine-reactive moieties, depending on your assay. Our sDIBO alkynes are improved versions of DIBO cyclooctynes, yielding conjugates that are less 'sticky' and emit lower signal background in biological samples.
Specifications
Specifications
| Quantity | for 5 mg of Ab |
| Kit Contents | Contains 1 vial of SiteClick Alexa Fluor 647 sDIBO Alkyne for Antibody Labeling in 250 μL DMSO and 1 antibody concentrator |
| Filter Type | Cy5 |
| Content And Storage | Store in refrigerator (2°C to 8°C) and protect from light. |
| Chemical Reactivity | Azide |
| Flow Cytometer Laser Lines | Red |
| Labeling Method | Click Chemistry |
| Labeling Scale | 5 mg |
| Shipping Condition | Wet Ice |
| Reactive Moiety | sDIBO Alkyne |
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Frequently Asked Questions (FAQs)
Structurally, they are the same. The only difference is in the manner of packaging. SiteClick Alexa Fluor 647 sDIBO Alkyne is packaged for labeling 1 to 5 mg of antibody per the SiteClick labeling protocol with the actual amount being proprietary. Click-iT Alexa Fluor 647 sDIBO Alkyne is packaged as a 0.5 mg of reagent for use with any copper-free click reaction.
The protocol is very simple. Expose the azide-modified sample to the sDIBO alkyne (final concentration in the range of 1 to 5 µM is a reasonable starting range) in aqueous buffer of media for 1-2 hrs. Optimize the final working concentration and incubation time per the sample/application. The azide-modified target must have the azide accessible for reacting with the sDIBO, and the labeling solution should not include any sodium azide.
Both the original DIBO alkynes and the sDIBO alkynes utilize the same fluorescent dyes or biotin for detection. The sDIBO alkynes have improved solubility in aqueous solutions and less propensity to bind non-specifically compared to the original DIBO alkynes.
The labeling should work with the buffer that your antibody is in. The sodium azide and glycerol will be removed in the buffer exchange with the purification column. This step also concentrates the antibody. The spin column has a 50 kDa MW cut-off, so it will also concentrate the BSA. However, this shouldn't be a problem as our in-house testing has shown that the SiteClick assay can label antibodies in the presence of BSA.
For Research Use Only. Not for use in diagnostic procedures.
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