AAT Bioquest

Protonex™Green 500-Latex Bead Conjugate

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Spectral properties
Extinction coefficient (cm -1 M -1)4000
Excitation (nm)445
Emission (nm)503
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageRefrigerated (2-8 °C); Minimize light exposure


Extinction coefficient (cm -1 M -1)
Excitation (nm)
Emission (nm)
Protonex™ Green 500-Latex Bead Conjugates exhibit distinctive pH-dependent fluorescence characteristics, becoming significantly more fluorescent in acidic conditions, unlike most conventional fluorescent dyes that increase in fluorescence at higher pH levels. This unique feature makes them highly suitable for investigating phagocytosis and its modulation by various drugs or environmental factors. The beads show minimal fluorescence outside of cells, eliminating the need for wash steps and enhancing their suitability for live-cell imaging. Protonex™ Green 500-Latex Bead Conjugates are particularly effective in highlighting acidic cellular compartments such as phagosomes, lysosomes, and endosomes, where they emit bright green fluorescence. Additionally, these conjugates can be used in combination with red fluorescent dyes like RFP, Calbryte™ 630 calcium dye, calcein red, or Cy5-labeled antibodies for multiplexed cell functional analysis. Their spectral properties are similar to those of FITC, allowing for the use of common FITC filter sets in assays involving Protonex™ Green 500-Latex Bead Conjugates, further facilitating their integration into existing experimental setups.


Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom

Example protocol


Chemical and Physical Properties
Solids Content:
1% in PBS
Number of Microspheres per mL:
445/503 nm
Mean Diameter:
0.72 µm



The following is a recommended protocol for granulocytes. This protocol only provides a guideline and should be modified
according to your specific experimental conditions. 

  1. Prepare cells as desired. For example, prepare the granulocytes at 107 cells/mL with Hanks and 20 mM Hepes buffer (HHBS), and add 100 μL to a polypropylene tube.

    Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density. 

  2. Add 1-10 μL of the Protonex™ Green 500-Latex Bead Conjugate to the tube and incubate with gentle shaking for 30 minutes at 37˚C.

    Note: Each cell line should be evaluated on an individual basis to determine the optimal incubation time. 

  3. Prepare an identical sample that is incubated at 4˚C and label it as a control.

  4. At the end of the 30-minute incubation, stop phagocytosis by adding 2mL of ice-cold HHBS and mix well.

  5. Wash the cells 2 times with cold HBSS. 

  6. Resuspend the cells in 500 μL of cold HBSS, keep the samples at 4˚C, and analyze immediately using a fluorescence microscope equipped with a FITC filter set.

    Note: For fluorescence microplate readers, monitor the fluorescence intensity at Ex/Em = 450/510 nm (Cutoff = 490 nm).


Open in Advanced Spectrum Viewer

Spectral properties

Extinction coefficient (cm -1 M -1)4000
Excitation (nm)445
Emission (nm)503


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Rapamycin treatment induces tubular proteinuria: role of megalin-mediated protein reabsorption.
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