Fluorescent Hypochlorite Detection Kit

1 Citation

Two cell-permeable dyes hydroxyphenyl fluorescein (HPF) and aminophenyl fluorescein (APF) detect hypochlorite production in cells. Hypochlorite production is observed by increased fluorescence of APF loaded cells with no increase in HPF loaded cells

Figure 1. Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer (as described in step V 1. above) and seeded in glass bottom dishes. The cells were then loaded with APF or HPF (10 mM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL).
Figure 2. Fluorescence images were acquired before and 10 minutes after stimulation. Excitation: 488 nm emission: 505-550 nm barrier filters. Hypochlorite production can be detected with increase APF fluorescence and no increase in HPF fluorescence.
Figure 1. Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer (as described in step V 1. above) and seeded in glass bottom dishes. The cells were then loaded with APF or HPF (10 mM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL).
Figure 1. Detection of Hypochlorite ( -OCl ) in neutrophils. Neutrophils were isolated from porcine blood, washed in Krebs-Ringers phosphate buffer (as described in step V 1. above) and seeded in glass bottom dishes. The cells were then loaded with APF or HPF (10 mM final) by incubation for 30 minutes at room temperature. The Dye-loaded neutrophils were stimulated with PMA (2ng/mL).
Figure 2. Fluorescence images were acquired before and 10 minutes after stimulation. Excitation: 488 nm emission: 505-550 nm barrier filters. Hypochlorite production can be detected with increase APF fluorescence and no increase in HPF fluorescence.

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Data Sheet

SKU: FLOCL100-2

Size: 100 Tests
Price:
$506
Hydroxyphenyl fluorescein (HPF) and Aminophenyl fluorescein (APF) are selective for the detection of highly reactive oxygen species (hROS). HPF has little reactivity towards other ROS such as: singlet oxygen (O), superoxide (O2– •), hydrogen peroxide (H2O2), nitric oxide (NO•), and alkyl peroxide (RO2•). HPF/APF are cell permeable and can be used in combination to detect hypo- chlorite (-OCl) production in cells. Hypochlorite can be detected by loading two samples, one with APF and the other with HPF. Hypochlorite production is visualized by increase in fluorescence of APF loaded cells and no increase in fluorescence in HPF loaded cells.
Flow Cytometer, Fluorescence plate reader, Fluorescence Microscope
2-8°C
Ships overnight (domestic), International Priority Shipping, NOT FOR SALE IN JAPAN.
United States
  • Part# 4012: HPF Dye, 1 Vial (2-8C)
  • Part# 4011: APF Dye, 1 Vial (2-8C)

    • Product Specific References

    PMID Publication
    30349873O'Grady, K. P., et al. 2018. Drug-Free ROS Sponge Polymeric Microspheres Reduce Tissue Damage from Ischemic and Mechanical Injury. ACS Biomater Sci Eng, 1251-1264.

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