Average mol wt 4000 fitcglucose 1250 describes a defined grade of fluorescent glucose conjugate where the average molar mass centers near 4000 Da and the fluorescein isothiocyanate substitution ratio is 1250. This labeling density supports bright detection in enzymatic assays and flow applications while maintaining reasonable solubility.
The balance between molecular weight and labeling efficiency makes this product suitable for glucose uptake studies, transport assays, and quantitative binding experiments in physiological buffer systems.
| Attribute | Value | Unit | Notes |
|---|---|---|---|
| Average molecular weight | 4000 | Da | Target specification for batch release |
| FITC labeling ratio | 1250 | mol glucose/mol FITC | Indicates molar ratio of dye to glucose |
| Impurity profile | <5 | % free FITC | Measured by chromatography |
| Storage condition | -20 | °C | Lyophilized powder recommended |
Physicochemical properties of average mol wt 4000 fitcglucose 1250
Molecular weight and labeling efficiency
At average mol wt 4000 fitcglucose 1250, the conjugate maintains a defined size that simplifies stoichiometric calculations in kinetic models. The labeling ratio of 1250 ensures sufficient fluorophore density for sensitive detection without overly perturbing glucose conformation.
Purity and stability considerations
Analytical HPLC confirms high chemical purity, with controlled free FITC content to minimize background fluorescence. Proper lyophilization and storage at -20 °C preserve activity over extended periods, supporting reproducible results across experiments.
Application scenarios for average mol wt 4000 fitcglucose 1250
Enzyme activity and transport assays
This product is commonly used in hexokinase and glucose permease assays, where fluorescence-based readouts enable real-time monitoring of enzymatic conversion and membrane translocation under defined conditions.
Flow cytometry and imaging protocols
In cellular imaging and flow cytometry, the labeled glucose provides bright, specific signal when combined with appropriate filters, allowing researchers to track glucose uptake in heterogeneous cell populations with minimal cross-talk.
Experimental considerations and handling
Buffer compatibility and quenching strategies
Formulation in mildly alkaline or neutral buffers preserves FITC integrity while minimizing nonspecific adsorption. Including inert protein stabilizers can further reduce surface-bound artifacts and improve quantitative accuracy in downstream measurements.
Calibration and standard curve design
Constructing calibration curves with known concentrations of average mol wt 4000 fitcglucose 1250 ensures accurate quantitation of enzymatic activity or transporter expression. Matching excitation and emission settings to the fluorophore profile maximizes signal fidelity and reduces measurement error.
Best practices and recommendations for average mol wt 4000 fitcglucose 1250
- Verify molar ratio and purity by HPLC before experimental use
- Use appropriate excitation/emission filters to maximize signal-to-noise
- Include controls with unlabeled glucose to assess specificity
- Optimize incubation duration to remain within the linear detection range
- Store lyophilized material at -20 °C and protect from repeated freeze-thaw cycles
FAQ
Reader questions
Can this conjugate be used in high-throughput screening formats?
Yes, its defined properties and strong fluorescence support automation-compatible protocols, enabling efficient plate-based assays with minimal rework.
How should I determine the optimal incubation time for glucose uptake experiments?
Run preliminary time-course assays using fixed cell numbers and this conjugate, then select an interval within the linear range to ensure proportional signal relative to transport activity.
What wavelength settings are recommended for detection?
Excite near 490 nm and monitor emission around 520 nm to capture maximal fluorescein fluorescence while reducing instrument background and assay interference.
Is this material suitable for studies involving membrane-permeable glucose analogs?
Yes, its moderate size and labeling pattern allow reasonable membrane passage, making it compatible with assays that compare permeable and impermeable glucose species under controlled conditions.