2. Others Immunology/Inflammation NF-κB Metabolic Enzyme/Protease
  3. Fluorescent Dye Reactive Oxygen Species (ROS)
  4. Hydroxyphenyl Fluorescein

Hydroxyphenyl Fluorescein  (Synonyms: HPF; 3'-p-(Hydroxyphenyl) fluorescein)

Cat. No.: HY-111330 Purity: 99.91%
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Hydroxyphenyl Fluorescein (HPF) is a stable ROS fluorescent probe dye. Hydroxyphenyl Fluorescein has stronger specificity and stability than H2DCFDA (HY-D0940). Hydroxyphenyl Fluorescein can produce strong green fluorescence through hydroxyl radical reaction with intracellular peroxynitroso. Hydroxyphenyl Fluorescein can be applied for fluorescence microscopy, high-throughput imager, luciferase microplate reader or flow cytometry. Ex/Em=490/515 nm.

For research use only. We do not sell to patients.

CAS No. : 359010-69-8

Size Price Stock Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO
ready for reconstitution
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10 mM * 1 mL in DMSO In-stock
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Customer Review

Based on 11 publication(s) in Google Scholar

Other Forms of Hydroxyphenyl Fluorescein:

Hydroxyphenyl Fluorescein Related Antibodies

Top Publications Citing Use of Products

    Hydroxyphenyl Fluorescein purchased from MedChemExpress. Usage Cited in: Adv Mater. 2025 Jan 10:e2415030.  [Abstract]

    CLSM imaging of ·OH in 4T1 cells following treatment with various nanoreactors. Scale bar: 20 µm.
    Intracellular ·OH levels were determined using Hydroxyphenyl Fluorescein (HPF, 10 µM; 30 min). The results demonstrated a significant increase in intracellular ·OH levels after treatment with F/G/H-GNC.

    Hydroxyphenyl Fluorescein purchased from MedChemExpress. Usage Cited in: Theranostics. 2025 Jun 9;15(14):6665-6685.  [Abstract]

    Detection of intracellular O2•- and •OH using DHR123 (20 μM) and Hydroxyphenyl Fluorescein (HPF, 20 μM) indicators in the presence of T-T NPs, with or without light exposure under hypoxia.

    Hydroxyphenyl Fluorescein purchased from MedChemExpress. Usage Cited in: Eur J Med Chem. 2025 Nov 15:298:118016.  [Abstract]

    Compare ROS generation of MeTS2IC with Ce6:·OH analyzed by Hydroxyphenyl Fluorescein (HPF, 5 μM) under dark and white light irradiation (10 mW/cm2).

    Hydroxyphenyl Fluorescein purchased from MedChemExpress. Usage Cited in: ACS Appl Mater Interfaces. 2024 Oct 2;16(39):52068-52079.  [Abstract]

    Confocal imaging planes and 3D images of Hydroxyphenyl Fluorescein (HPF, 10 μM; 30 min) staining after the biofilms were incubated with Ru4 (10 μM) for 9 h.

    • Biological Activity

    • Purity & Documentation

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    • Customer Review

    Description

    Hydroxyphenyl Fluorescein (HPF) is a stable ROS fluorescent probe dye. Hydroxyphenyl Fluorescein has stronger specificity and stability than H2DCFDA (HY-D0940). Hydroxyphenyl Fluorescein can produce strong green fluorescence through hydroxyl radical reaction with intracellular peroxynitroso. Hydroxyphenyl Fluorescein can be applied for fluorescence microscopy, high-throughput imager, luciferase microplate reader or flow cytometry. Ex/Em=490/515 nm[1].

    In Vitro

    Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
    1. Preparation of HPF working solution
    1.1 Preparation of the stock solution
    Dissolve HPF in DMSO to obtain 10 mM of HPF.
    1.2 Preparation of HPF working solution
    Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of HPF working solution.
    Note: Please adjust the concentration of HPF working solution according to the actual situation.
    2. Cell staining
    2.1 Cell preparation.
    For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
    For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
    2.2 Add 1 mL of HPF working solution, and then incubate at room temperature for 30 minutes.
    2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
    2.4 Wash twice with PBS, 5 minutes each time.
    2.5 Resuspend cells with 1 mL serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    Hydroxyphenyl Fluorescein Related Antibodies
    Molecular Weight

    424.40

    Formula

    C26H16O6
    CAS No.
    Appearance

    Solid

    Color

    Yellow to orange

    Emission (Em)

    515
    Excitation (Ex)

    490
    SMILES

    OC1=CC=C(OC2=CC3=C(C4(C5=CC=C(C=C5O3)O)C6=C(C(O4)=O)C=CC=C6)C=C2)C=C1
    Shipping

    Room temperature in continental US; may vary elsewhere.
    Storage

    -20°C, protect from light

    *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 125 mg/mL (294.53 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 2.3563 mL 11.7813 mL 23.5627 mL
    5 mM 0.4713 mL 2.3563 mL 4.7125 mL
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

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    Volume (start)

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    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Calculation results:
    Working solution concentration: mg/mL
    Purity & Documentation
    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 2.3563 mL 11.7813 mL 23.5627 mL 58.9067 mL
    5 mM 0.4713 mL 2.3563 mL 4.7125 mL 11.7813 mL
    10 mM 0.2356 mL 1.1781 mL 2.3563 mL 5.8907 mL
    15 mM 0.1571 mL 0.7854 mL 1.5708 mL 3.9271 mL
    20 mM 0.1178 mL 0.5891 mL 1.1781 mL 2.9453 mL
    25 mM 0.0943 mL 0.4713 mL 0.9425 mL 2.3563 mL
    30 mM 0.0785 mL 0.3927 mL 0.7854 mL 1.9636 mL
    40 mM 0.0589 mL 0.2945 mL 0.5891 mL 1.4727 mL
    50 mM 0.0471 mL 0.2356 mL 0.4713 mL 1.1781 mL
    60 mM 0.0393 mL 0.1964 mL 0.3927 mL 0.9818 mL
    80 mM 0.0295 mL 0.1473 mL 0.2945 mL 0.7363 mL
    100 mM 0.0236 mL 0.1178 mL 0.2356 mL 0.5891 mL
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    Hydroxyphenyl Fluorescein Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

    Keywords:

    Hydroxyphenyl Fluorescein359010-69-8HPF 3'-p-(Hydroxyphenyl) fluoresceinFluorescent DyeReactive Oxygen Species (ROS)ROSprobeOHindicatorfluoresceperoxynitriteInhibitorinhibitorinhibit

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