ELISA
Coat the plate with immunizing antigen. Blue line:Immune serum; Orange line: Pre-immune serum.
WB
Western blot analysis using LPAR4 antibody against Hela(1),Cos-7(2),K562(3),B16(4),Hepg2(5),U251(6),A549(7),Nih3t3(8) cell,Mouse brain tissue(9),Mouse liver tissue(10),Mouse lung tissue(11),Rat kidney tissue(12),Rat spleen tissue(13),Rat testes tissue(14) .12% SDS-PAGE gel. Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane, and block it with TBST plus skimmed milk powder for one hour. Dilute the primary antibody with the antibody diluent at a ratio of 1:1000, and incubate it overnight at 4°C. Wash the membrane three times with TBST, 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP, at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST again, 5 minutes each time. Use ECL for luminescence. staining time: 60S.