Mix-n-Stain™ antibody labeling kits are a revolutionary antibody labeling technology that allows you to label your antibody with one of our superior CF® dyes in 30 minutes without a purification step. The labeling procedure tolerates many common buffer components and antibody stabilizers. Now with an extended shelf life of at least 1 year.
For CF® dye, cyanine dye, FITC, biotin, DNP, and dig kits, see the Mix-n-Stain™ Kit Compatibility and Protocol Selection Flowchart, below. We also recommend downloading the updated Product Protocol and completing the pre-labeling checklist to find the right protocol and kit size to use for each antibody you wish to label. Learn more about CF® dyes on our CF® Dye Technology Page, or download the CF® Dye Brochure.
CF® dyes are highly water soluble, small organic dyes designed by scientists at Biotium for labeling proteins and nucleic acids. With a series of over 20 colors (and growing), many of our CF® dyes are brighter and more photostable than competing dyes. For more information please see the product flyers for individual CF® dyes, the CF® Dye Selection Guide, and our CF® Dye FAQs.
CF® initially was an abbreviation for Cyanine-based Fluorescent dyes. These were the first patented CF® dyes based on cyanine dye structures. Since then, our CF® dye patent portfolio has expanded to include four different fluorescent dye core structures that cover the fluorescence spectrum from UV to NIR.
The exact chemical structures of CF® dyes are currently confidential but will be fully disclosed at a later stage when pending patents become granted. In general terms, the structure of a CF® dye may be divided into two parts: a) dye core structure (i.e. the aromatic ring skeleton that defines the dye’s color or absorption/emission wavelengths), and b) core structure-modifying elements. At present, CF® dyes bear the core structures of coumarin, pyrene, rhodamine or cyanine dyes. Blue fluorescent CF® dyes are based on coumarin or pyrene dye core structure, while green to near-IR CF® dyes are based on either cyanine or rhodamine dye core structures. Core structure-modifying elements refer to various chemical attachments to the core structure and are a key aspect of the CF® dye invention that makes CF® dyes superior to other commercial dyes.
There is no simple answer to this question as the quantum yield of a fluorescent dye can vary widely, depending on the dye’s micro-environment. For example, the quantum yield of a dye attached to a protein may be very different from the quantum yield of the free dye. For dyes attached to a protein, the quantum yield is highly dependent on how many molecules of the dye are attached to the protein (i.e. degree of protein labeling). In general, a higher degree of protein labeling leads to a lower dye quantum yield due to fluorescence quenching via dye-to-dye interaction. For this reason, as the degree of labeling increases, fluorescence intensity of the labeled protein will eventually reach a maximum and start to decline thereafter. In fact, one of the best ways to compare the relative quantum yields of different dyes is to plot the total fluorescence of the labeled proteins as a function of degree of labeling by the dyes as we have done with CF® dyes and other commercial dyes. CF® dyes generally give higher slopes than other commercial dyes in the plots, suggesting less quantum yield decline with increasing degree of protein labeling.
Mix-n-Stain™ CF® Dye Antibody Labeling Kits allow you to covalently label ≤5 ug up to 100 ug of your antibody in just 30 minutes, with minimal hands-on time and no purification. The kits tolerate common antibody buffer formulations. With a slight modification in protocol, antibodies even can be labeled in the presence of BSA, gelatin, or ascites fluid, for unrivaled convenience. Because CF® dyes are highly water soluble, the presence of other proteins like BSA or gelatin in the labeling reaction has minimal effect on background fluorescence, because any labeled non-antibody proteins readily wash away during immunofluorescence staining.
These kits feature a choice of 29 of Biotium’s next-generation CF® dyes, which were designed for advantages in brightness and photostability compared to Alexa Fluor®, DyLight®, and other fluorescent dyes. Learn more about CF® Dyes.
Label 5-20 ug, 20-50 ug, or 50-100 ug Ab
Choice of 29 CF® Dye colors
Less than 30 seconds of hands-on time
30 minutes total reaction time
No purification of the product is required, for 100% recovery
Compatible with common antibody stabilizers
Choose the Right Labeling Kit for Your Antibody
Mix-n-Stain™ Antibody Labeling Kits are very simple to use (see the workflow overview above). But before you begin, you must check that your antibody meets the compatibility requirements for labeling, and choose the right labeling protocol. Download the updated and expanded Product Protocol for a labeling check list to help you select the right kit size and labeling protocol for your antibody. Also see our Kit Compatibility and Protocol Selection Flowchart.
See the product table below for a quick reference of the maximum excitation and emission of each CF® dye, or see our CF® Dye Brochure for more detailed dye information.
CoverGrip™ Coverslip Sealant is the first product specifically designed for sealing the edges of wet-mounted coverslips for fluorescence microscopy. Unlike nail polish, CoverGrip™ contains no ingredients that can leach into aqueous mounting medium and affect specimen fluorescence.
Easily label your antibody with phycobiliproteins APC, R-PE and PerCP, or tandem dyes APC-CF®750T and RPE-CF®647T, in about 4 hours. The labeling procedure tolerates many common buffer components and antibody stabilizers.
Paraformaldehyde, 4% in PBS is a ready-to-use fixation solution for cells or tissues. It is electron microscopy-grade paraformaldehyde dissolved in pH 7.4 PBS, and is stabilized by packaging under argon with no methanol added.
TrueBlack® Lipofuscin Autofluorescence Quencher, 20X in DMF, is a new reagent for quenching lipofuscin autofluorescence in tissue sections for immunofluorescence staining. It can also help reduce autofluorescence from other sources like extracellular matrix.