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Protonex™Green 500-Latex Bead Conjugate

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.

Example protocol

AT A GLANCE

Chemical and Physical Properties
Solvent:
Water
Solids Content:
1% in PBS
Number of Microspheres per mL:
~4e+10
Ex/Em:
445/503 nm
Mean Diameter:
0.72 µm

SAMPLE EXPERIMENTAL PROTOCOL

Important

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

Protocol
  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).

Spectrum

References

View all 50 references: Citation Explorer
Modified Danzhi Xiaoyao Powder (MDXP) improves the corneal damage in dry eye disease (DED) mice through phagocytosis.
Authors: Liu, Pei and Jiang, Pengfei and Yu, Yunfeng and Tan, Kang and Qin, Gen-Yan and Liu, Tingting and Tian, Sainan and Peng, Jun and Peng, Qinghua
Journal: Journal of ethnopharmacology (2024): 117544
Role of Inhibiting Inflammation of LC3-Associated Phagocytosis in Dry Eye Disease.
Authors: Zhang, Sasa and Liu, Xing and Li, Cui and Wang, Qian and Yang, Shanshan and Peng, Xudong and Hu, Liting and Zhao, Guiqiu and Lin, Jing
Journal: Current eye research (2024): 25-32
Amphiphilic Cell-Penetrating Peptides Containing Arginine and Hydrophobic Residues as Protein Delivery Agents.
Authors: Moreno, Jonathan and Zoghebi, Khalid and Salehi, David and Kim, Lois and Shoushtari, Sorour Khayyatnejad and Tiwari, Rakesh K and Parang, Keykavous
Journal: Pharmaceuticals (Basel, Switzerland) (2023)
Sessile Trichomes Play Major Roles in Prey Digestion and Absorption, While Stalked Trichomes Function in Prey Predation in Byblis guehoi.
Authors: Li, You-Xian and Chen, Alvin and Leu, Wei-Ming
Journal: International journal of molecular sciences (2023)
Mouse embryonic stem cell-derived blood-brain barrier model: applicability to studying antibody triggered receptor mediated transcytosis.
Authors: Jezierski, Anna and Huang, Jez and Haqqani, Arsalan S and Haukenfrers, Julie and Liu, Ziying and Baumann, Ewa and Sodja, Caroline and Charlebois, Claudie and Delaney, Christie E and Star, Alexandra T and Liu, Qing and Stanimirovic, Danica B
Journal: Fluids and barriers of the CNS (2023): 36
Page updated on December 6, 2024

Ordering information

Price
Unit size
Catalog Number21223
Quantity
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Additional ordering information

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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

Storage

Refrigerated (2-8 °C); Minimize light exposure
UNSPSC12352200

Platform

Fluorescence microscope

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