Preparation and identification of polyclonal antibody against cysteinyl leukotriene receptor 2.
- Author:
Li-Ping ZHANG
1
;
Chun-Zhen ZHAO
;
Wen-Zhen SHI
;
Ling-Ling QI
;
Yun-Bi LU
;
Yong-Mei ZHANG
;
Li-Hui ZHANG
;
San-Hua FANG
;
Jian-Fang BAO
;
Jian-Gen SHEN
;
Er-Qing WEI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; immunology; Brain; metabolism; Kidney; metabolism; Lung; metabolism; Mice; Rabbits; Rats; Rats, Sprague-Dawley; Receptors, Leukotriene; immunology; metabolism
- From: Journal of Zhejiang University. Medical sciences 2009;38(6):591-597
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare and identify a polyclonal antibody against cysteinyl leukotriene receptor (CysLT(2)receptor).
METHODSRabbits were immunized with KLH-coupled CysLT(2) receptor peptide to prepare the polyclonal antibody (pAb). The titer of the pAb in rabbit plasma was detected by indirect ELISA, and the specificity of the pAb was tested by antigen blockade. The tissue distribution of CysLT(2) receptor was detected by Western blot and immunohistochemistry with the prepared pAb.
RESULTThe pAb showed a titer higher than 1/1047296, and was specific to CysLT(2) receptor, without cross-reaction with the antigens of CysLT(1) receptor and GPR17. A higher expression of CysLT(2) receptor in kidney, brain and lung of rats and mice was detected by Western blot analysis using the prepared pAb. The molecular weight of CysLT(2) receptor protein was about 40 kD. Immunohistochemical examination showed that CysLT(2) receptor was expressed mainly in the neuron, and partly in astrocytes in rat brain.
CONCLUSIONThe prepared CysLT(2) receptor pAb has high sensitivity and specificity, and can be used in Western blot and immunohistochemistry.
