[Preparation and identification of polyclonal antibody against cysteinyl leukotriene receptor 2].
To prepare and identify a polyclonal antibody against cysteinyl leukotriene receptor (CysLT(2)receptor).



Rabbits 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.



The 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.



The prepared CysLT(2) receptor pAb has high sensitivity and specificity, and can be used in Western blot and immunohistochemistry.
Zhang
Li-Ping
LP
Department of Pharmacology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
Zhao
Chun-Zhen
CZ
Shi
Wen-Zhen
WZ
Qi
Ling-Ling
LL
Lu
Yun-Bi
YB
Zhang
Yong-Mei
YM
Zhang
Li-Hui
LH
Fang
San-Hua
SH
Bao
Jian-Fang
JF
Shen
Jian-Gen
JG
Wei
Er-Qing
EQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D001921:Brain / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D018077:Receptors, Leukotriene / Q000276:immunology* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D018077:Receptors, Leukotriene; C412375:cysteinyl leukotriene receptor 2