We offer a variety of anti-fluorescent dye antibodies that recognize specific fluorophores and, in most cases, quench their fluorescence. These anti-dye antibodies, including those that recognize the tetramethylrhodamine (TRITC) and Texas Red™ fluorophores, can also serve as cell-impermeant probes for determining whether fluorescent dye-conjugated ligands, proteins, bacteria, or other biomolecules have been internalized by endocytic or pinocytic processes. Our anti-tetramethylrhodamine (A6397) and anti-Texas Red (A6399) antibodies have been raised against the tetramethylrhodamine and Texas Red fluorophores, respectively. These antibodies quench much of the fluorescence of the complementary dyes. However, due to the related chemical structures, the anti-tetramethylrhodamine antibody crossreacts with the Texas Red fluorophore and the anti-Texas Red antibody cross-reacts with tetramethylrhodamine. Also, both antibodies tightly bind the Rhodamine Red™ fluorophore. Thus, either antibody can serve as an effective probe for tetramethylrhodamine, Texas Red, or Rhodamine Red dyes.
We use a sensitive quenching assay to ensure that this antibody is provided at a consistently high titer value. As supplied, 20 µL of the antibody solution is certified to produce ≥50% of the maximal fluorescence quenching of 1 mL of a 50 nM solution of tetramethylrhodamine, assayed in 100 mM sodium phosphate, pH 8.0. Maximal quenching for tetramethylrhodamine is ~80% of the fluorescence of the free dye. Due to steric hindrance, maximal fluorescence quenching of the tetramethylrhodamine covalently bound to protein may be significantly less.
Tetramethylrhodamine (TRITC) is a bright orange-fluorescent traditional dye with excitation ideally suited to the 532 nm laser line. It has been used extensively to label protein conjugates such as phalloidins for cytoskeleton counterstaining or antibody conjugates for immunofluorescence and other cellular imaging applications. Unlike biotin, which is an endogenous ligand in mitochondria, dye-based haptens permit background-free staining of cells and tissues and a sensitive alternative to biotin-streptavidin methods.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: Tetramethylrhodamine isothiocyanate