[Preparation and identification of a polyclonal antibody against novel cysteinyl leukotriene receptor GPR17].
OBJECTIVE
To prepare and identify a polyclonal antibody (pAb) against GPR17, a novel cysteinyl leukotriene receptor.


METHODS
Rabbits were immunized with KLH-coupled GPR17 peptide to prepare the 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. GPR17 tissue distribution was detected by Western blot with the pAb.


RESULTS
The pAb showed a titer as high as 1:16 364,and was not cross-reacted with the antigens of CysLT(1) and CysLT(2) receptors. A higher expression of GPR17 in the rat brain and heart was detected using the newly prepared pAb. The molecular weigh of GPR17 protein was about 43 kD.


CONCLUSIONS
The prepared GPR17 pAb has high sensitivity and specificity,and can be used in Western blot for detecting GPR17.
QI
Ling-ling
LL
Department of Pharmacology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
LU
Yun-bi
YB
SHI
Wen-zhen
WZ
ZHAO
Chun-zhen
CZ
ZHANG
Yong-mei
YM
CHEN
Li-ping
LP
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*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D011817:Rabbits; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000276:immunology*; D018077:Receptors, Leukotriene / Q000276:immunology*
D000911:Antibodies, Monoclonal; C121670:GPR17 protein, human; D043562:Receptors, G-Protein-Coupled; D018077:Receptors, Leukotriene; C083169:leukotriene D4 receptor