TRITC (tetramethylrhodamine isothiocyanate) conjugated anti-mouse primary antibodies are directly labeled immunoreagents that combine a mouse antigen-specific immunoglobulin with the orange-red emitting TRITC fluorophore. Unlike TRITC-conjugated secondary antibodies, which recognize mouse immunoglobulins, these primary antibodies bind their target mouse antigen directly, enabling single-step staining without the need for a secondary reagent.

Introduction

TRITC is a rhodamine-class fluorescent dye with an excitation maximum of approximately 550 nm and an emission maximum of approximately 570 nm. It is optimally excited by the 532 nm or 561 nm laser lines and detected using a 582/15 nm or 585/42 nm bandpass filter. TRITC exhibits good photostability and low pH sensitivity, making it suitable for both flow cytometry and imaging applications. Direct TRITC conjugates of anti-mouse primary antibodies are particularly valuable when a secondary antibody step is undesirable—for example, in mouse-on-mouse applications where endogenous immunoglobulins would produce high background, or in multiplexed panels where secondary reagents must be avoided.

Applications

• Flow cytometry: Direct immunophenotyping of mouse leukocyte subsets when a 532 nm or 561 nm laser is available; useful for multicolor panels alongside FITC, PE, and APC.
• Immunofluorescence microscopy: Sensitive detection of mouse antigens in cultured cells and tissue sections, particularly for dual-labeling with green fluorophores.
• Immunohistochemistry: Staining of frozen and formalin-fixed, paraffin-embedded mouse tissues.
• Western blotting: Fluorescent detection of mouse proteins on membranes.