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Mix-n-Stain CF™ Dye Antibody Labeling Kits

Mix-n-Stain™ antibody labeling kits are a revolutionary technology that dramatically simplifies the process of antibody labeling. The kits enable a researcher to covalently label ≤5 ug up to 100 ug of an antibody with one of 20 super bright and photostable CF™ dyes, biotin, or FITC in only 30 minutes and 100% recovery of antibody after labeling (Figure 1). The prepared conjugates perform comparably to  pre-labeled antibodies from leading suppliers. Because the labeling is covalent, the conjugates are stable for long-term storage, and ideal for multi-color imaging (Figure 2).

Each Mix-n-Stain™ kit comes with everything you need to perform the conjugation. Simply mix your antibody with a vial of pre-measured CF™ dye, biotin, or FITC in the buffer provided, a step taking less than 30 seconds of hands-on time. After waiting for another 30 minutes, without a separation step, you will have an optimally labeled antibody conjugate ready to be used in any immunofluorescence staining experiment.

CF488A GAM min x rat, CF555 Mix-n-Stain, CF640R phalloidin-IHC
Figure 2. Frozen section of rat testis stained with Mix-n-Stain CF488A labeled mouse anti-tubulin (microtubules, green), Mix-n-Stain CF555 labeled mouse anti-ZO1 (tight junctions, red), and CF640R phalloidin (cat. no. 00050) (actin filaments, cyan).

One breakthrough feature of Mix-n-Stain™ kits is that antibody labeling can be conducted in the presence of virtually any amount of BSA, gelatin or non-antibody proteins in ascites fluid when the new, modified protocol is used. Azide and low concentrations of Tris are also tolerated; an ultrafiltration spin column is included in the kit to quickly and easily remove incompatible small molecules like glycerol, glycine, or high concentrations of Tris. As a result, the kits are compatible with most commercial antibody formulations without an antibody purification step prior to the conjugation process.

Another major advantage of the labeling kits is our superior CF™ dyes, many of which are brighter and more photostable than Alexa Fluor® dyes and other similar next-generation fluorescent dyes. With a collection of 20 CF™ dyes spanning the entire visible and near-infrared spectra, you have unprecedented freedom to label any primary antibody with a color of your choice or biotin, making multi-color detection simple and affordable.

Frequently Asked Questions and Answers:

Q: Is the labeling chemical or physical?

A: The labeling is chemical, which means the dye is covalently attached to the antibody. Covalent labeling ensures no dye transfer or diffusion in multi-color detection employing multiple antibodies labeled with different CF Mix-n-Stain kits.

Q: What are the advantages of Mix-n-Stain kits over Invitrogen’s Zenon® antibody labeling kits?

A: There are four major advantages: 1) Unlike Zenon, Mix-n-Stain labeling covalently attaches the dye to the antibody, eliminating dye transfer or diffusion between antibodies during multi-color staining; 2) Mix-n-Stain conjugates are stable for at least 6 months in storage buffer, whereas Zenon complexes must be used within 30 minutes; 3) Mix-n-Stain conjugates are less bulky because the dyes are directly linked to the antibody, unlike Zenon complexes which use antibody fragments; 4) No post-staining fixation is required with Mix-n-Stain; 5) Unlike Zenon, Mix-n-Stain labeling is not species-specific. Read more…