Mix-n-Stain™ Antibody Labeling Technology

Direct labeling of primary antibodies is useful for flow cytometry applications and when primary antibodies from different species are not available for multi-color immunofluorescence experiments. Typically, labeling antibodies with reactive dyes requires antibody buffer exchange, long incubation times, and time-consuming purification steps to remove unconjugated dye. Mix-n-Stain antibody labeling kits are a breakthrough technology for simple and rapid small scale antibody labeling from 5 to 100 µg.

Figure 1. Evaluation of CF™ Dye Mix-n-Stain labeled antibodies.
 

A. Flow cytometry analysis (on BD FACS Calibur) of Jurkat cells stained with CF™633 Mix-n-Stain labeled mouse anti-human CD3 antibodies. For reference, cells were stained with commercial Alexa Fluor® 647-mouse anti-human CD3. Bars represent the relative fluorescence values of the geometric means. Fluorescence was analyzed using a BD FACSCalibur flow cytometer.

B. HeLa cells were stained with β-tubulin IgM conjugated with CF™633 Mix-n-Stain (red), followed by CF™488A phalloidin (green) and DAPI (blue). Images were captured on a Zeiss 510 Meta confocal microscope.

Simple Antibody Labeling Protocol – 3 Easy Steps

  1. Mix antibody with dye and buffer – 30 seconds hands on time
  2. Incubate antibody/dye for 30 minutes
  3. Conjugate is ready to use – No purification necessary!

Advantages for using Mix-n-Stain Labeling Kits

  • No need to calculate how much dye you should use - just mix your antibody with the entire amount of dye provided
  • There is no purification of conjugate necessary
  • The labeling reaction can tolerate the presence of common stabilizers, such as sodium azide, Tris, and low levels of glycerol, BSA or gelatin
  • Choose from over 19 of the brightest and most photostable dyes commercially available

Materials

     

CF and Mix-n-Stain are trademarks of Biotium, Inc.