The world’s simplest labeling protocol: Mix and then Stain ~ it really is that simple!
Mix-n-Stain™ CF™ dye small ligand labeling kits are designed for rapid labeling of small (MW ~ 150 – 5,000) and relatively high affinity biological ligands (or substrates). Labeling takes about 30 minutes, without a final purification step. The ligands to be labeled must contain an aliphatic amine group that is not required for biological activity of the ligand. The amine group will form a covalent linkage with the reactive CF dye provided in the kit. For example, suitable ligands or substrates include SNAP-tag®, CLIP-tag™ and HaloTag® ligands with an aliphatic amine. Many other small ligands are also possible candidates if they meet the criteria described above.
Simply mix your small molecule ligand with the reaction buffer and the optimally formulated dye provided, and incubate for 30 minutes, followed by a brief 5 minute quenching step. No reactive Mix-n-Stain™ dye is available at the end of labeling; therefore the conjugate is ready for cell staining and imaging without further purification. Mix-n-Stain™ labeled ligands perform comparably to commercially-available pre-labeled ligands (Figure 1), at a fraction of the cost.
A major advantage of Mix-n-Stain™ kits is our superior CF™ dyes, many of which are brighter and more photostable than Alexa Fluor® dyes and other similar commercial dyes. Several of our CF™ dyes, such as CF™405M, CF™633, CF™640R, CF™647, and CF™680R, have been demonstrated to be ideal probes for super-resolution and single molecule imaging (see references here). With a choice of ten CF™ dyes for cell surface labeling and three CF™ dyes for intracellular labeling, you have significant freedom to label your small molecule ligand with colors ranging from blue to near-infrared, making multi-color detection simple and affordable (Figures 2, 3). See the product list at the bottom of this page for more information.
For live cell imaging, the Mix-n-Stain™ kits are ideal to generate fluorescent chemical tags that provide smaller, brighter, and more versatile alternatives to fluorescent proteins. Labeling of various chemical tags with Mix-n-Stain™ kits yielded highly efficient and specific labeling of the target proteins both on surface and inside of living cells. The Mix-n-Stain kits have also been proven compatible with fixed cell imaging (Figure 3) and gel-based assays (see the Mix-n-Stain™ Small Ligand Flyer for more details).
Mix-n-Stain™ small ligand labeling kits for cell surface labeling are available in your choice of 10 CF™ dye colors for labeling ligands for cell surface targets, and three CF™ dye colors for ligands that bind intracellular targets. Also see our Mix-n-Stain™ Antibody Labeling Kits and other Reactive Dyes and Biotin Reagents.
|Dye||Catalog no.||Spectrally similar dyes||Ex (nm)||Em (nm)||Staining|
|CF405M||92362||Pacific Blue, BD Horizon V450||408||452||Surface|
|CF488A||92350||FITC, Alexa Fluor® 488||490||515||Surface|
|CF568||92351||Rhodamine Red, Alexa Fluor® 568||562||583||Surface|
|CF594||92352||Texas Red®, Alexa Fluor® 594||593||614||Surface|
|CF633||92353||Cy®5, Alexa Fluor® 633||630||650||Surface|
|CF640R||92354||Cy®5, Alexa Fluor® 647||642||662||Surface|
|CF647||92359||Cy®5, Alexa Fluor® 647||650||665||Surface|
|CF660C||92360||Alexa Fluor® 660||667||685||Surface|
|CF680||92361||Cy®5.5, Alexa Fluor® 660, IRDye®680LT||681||698||Surface|
|CF680R||92355||Cy®5.5, Alexa Fluor® 660, IRDye®680LT||680||701||Surface|
|CF408||92356||Pacific Blue, BD Horizon V450||408||450||Intracellular|
|CF500||92357||FITC, Alexa Fluor® 488||500||510||Intracellular|
|CF540||92358||Tetramethylrhodamine (TAMRA), TRITC, Alexa Fluor® 546, Cy®3||540||570||Intracellular|
|CF555||92364||Alexa Fluor® 555, Cy®3, TRITC||555||585||Intracellular|
|CF650||92363||Cy®5, Alexa Fluor® 647||650||670||Intracellular|
Mix-n-Stain and CF dyes are covered by US and international patents, and are trademarks of Biotium, Inc. SNAP-tag and CLIP-tag are trademarks or registered trademarks of New England BioLabs; HaloTag is a registered trademark of Promega Corporation; Pacific Blue, Texas Red, and Alexa Fluor are trademarks or registered trademarks of Invitrogen Corp; BD Horizon is a trademark of BD Biosciences; Cy Dye is a registered trademark of GE Healthcare; IRDye is a registered trademark of LI-COR Biosciences.