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Updated: Jul 20, 2026
Proteomics to Identify Proteins Interacting with P2X2 Ligand-Gated Cation Channels
Published on: May 19, 2009
[Detection of P2z/P2x7 receptors]
Liming Peng1, Hanshuo Yang, Qin Zhang
1Clinical Laboratory, West China Hospital of Sichuan University, Chengdu 610041, China.
Objective:
To detect the P2z/P2x7 receptors of human leukocytes.
Methods:
Re-suspending different kinds of leukemic cells, including CEM, THP-1, HL-60, NB4, Dami/Damin and Raji cells in buffer; adjusting their concentration to 2 x 10(7)/ml, and then detecting the P2z/P2x7 receptors respectively.
Results:
(1) It was found that 0.4 mmol/L ATP enhanced obviously the fluorenscence strength of THP-1, NB4 and HL-60 cells, the activity rates were 54.91%, 39.45% and 30.46%, respectively. But for CEM, Dam and Raji cells, the activity rates were as low as 10.43%, 13.21% and 2.67%, respectively. KN-62 inhibited obviously the fluorescence strength of THP-1 cells, the inhibition rate reached 88.74%, but for NB4 and HL-60 cells the inhibition rates were only 15.81% and 12.50% respectively, and Raji cells were not affected by KN-62. (2) All the leukemic cells detected with polyclonal antibody were found with positive result.
Conclusion:
THP-1 cells have P2z/P2x7 receptors. The receptors in NB4 and HL-60 belong to P2 receptor family, but they are not P2z/P2x7 receptors. The polyclonal antibody used in immunohistochemical ABC method is not specific to P2z/P2x7 receptors.
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