Frequently asked questions about the Mix-n-Stain™ labeling kit for antibody conjugation with CF™ dyes.
This FAQ addresses common questions about the Mix-n-Stain™ Labeling Kit, including dye removal, multi-color staining, protein compatibility, sensitivity, and comparisons with other kits. It also explains how to select the correct kit size based on antibody quantity and provides guidance on antibody concentration and troubleshooting staining issues.
The kit uses covalently linked CF™ dyes for direct labeling, eliminating dye transfer and offering advantages such as stability, no optimization needed, and no post-staining fixation. It is optimized for IgG antibodies and includes a reference guide to ordering CF™ dyes with their excitation/emission wavelengths.
The unique formulations of our dyes and buffers and the labeling strategy have completely removed this concern, which normally has to be dealt with when using conventional antibody labeling methodology.
Mix-n-Stain™ labeling results in covalent linkage of dye and antibody, so there will be no dye diffusion or transfer.
Mix-n-Stain™ kits are optimized for labeling IgG antibodies. We do not recommend them for labeling other proteins. Mix-n-Stain™ labeling conditions may cause denaturation of IgM antibodies.
Mix-n-Stain™ antibody labeling kits use novel CF dyes which are brighter and more photostable than the dyes provided in Lightning Link kits. Mix-n-Stain™ kits are sized for labeling smaller amounts of antibody and are sold as a single labeling, providing more flexibility compared to Lightning Link kits.
For each CF dye, there are three labeling kits for labeling of antibody quantities in three different ranges: 1) 5-20 µg, 2) 20-50 µg, and 3) 50-100 µg. For antibody labeling in the absence of stabilizer protein, select a kit that matches with the amount of your antibody. For antibody labeling in the presence of stabilizer protein or ascites fluid, see Table 2 of the product protocol.
There is no need to measure the dye amount or vary the reaction time as long as the amount of your antibody to be labeled falls within the range specified for each kit. With Mix-n-Stain™ labeling kits optimal labeling is ensured because of the proprietary dyes and reaction buffer.
Check with the antibody manufacturer to confirm that the antibody formulation and concentration are compatible with the kit labeling protocol you selected. You should confirm that your primary antibody is sensitive and specific for your application using indirect labeling before attempting direct labeling. You may need to use a higher concentration of primary antibody to achieve similar signal intensity with direct labeling as with indirect labeling.