6-Carboxy-H2DCFDA | CAS 247044-02-6

6-Carboxy-H2DCFDA is a carboxylated, cell-trappable fluorogenic probe based on the H2DCFDA scaffold, designed for oxidative-stress and reactive oxygen species related workflows. After intracellular ester processing and oxidation, it forms a fluorescent product that is commonly read by fluorescence microscopy and plate-based assays. The carboxyl group supports downstream conjugation or retention strategies in imaging and materials-adjacent experiments where controlled probe localization is beneficial.

1. Oxidative Stress Imaging

6-Carboxy-H2DCFDA is used by cell biology and chemical biology groups to visualize oxidative stress dynamics in cultured cells using fluorescence microscopy. Researchers typically apply the probe in time-course experiments to compare relative oxidative burden across experimental conditions, including stimulation or antioxidant treatment controls, while monitoring the resulting fluorescence signal in adherent or suspension formats compatible with standard imaging workflows. The probe's fluorogenic response enables signal generation primarily after cellular processing and oxidation, supporting studies focused on ROS-associated changes rather than direct dye labeling.

2. Plate-Based ROS Assays

6-Carboxy-H2DCFDA is frequently incorporated into microplate fluorescence assays for high-throughput screening of oxidative stress modulation in cell-based formats. Assay developers use the probe to generate a readout that scales with ROS-related oxidation of the probe, enabling comparative quantification across multiple conditions in 96- or 384-well plates. This workflow is commonly used in reagent optimization, pathway perturbation studies, and screening campaigns where a robust fluorescence endpoint is preferred for automation-compatible data collection.

3. Flow Cytometry ROS Readout

6-Carboxy-H2DCFDA supports flow cytometry-based assessment of oxidative stress at the single-cell level, where researchers quantify fluorescence intensity distributions across cell populations. In typical cytometry workflows, the probe is loaded into cells and the resulting oxidized fluorescence is measured to resolve heterogeneity in oxidative status that may be masked by bulk measurements. The carboxylated variant is also leveraged in experimental designs that benefit from improved retention or downstream handling relative to non-carboxylated analogs, aiding consistent comparisons between treatment groups.

4. Probe-Conjugate Development

6-Carboxy-H2DCFDA is used as a starting fluorogenic handle for building ROS-responsive labeling tools and conjugate-adjacent research reagents. Chemical biology groups may exploit the carboxyl functionality to couple the probe to linkers or biomaterial surfaces for experiments where probe immobilization, localized reporting, or tailored probe presentation is desired. This application is particularly relevant for developing experimental platforms that combine oxidative readouts with controlled spatial context, such as surface-associated assays or imaging formats where probe localization affects interpretability.