Immuno-blot detection of hemangiopoietin in the human fetal liver.
- Author:
Ying-Song QIN
1
;
Zhong-Chao HAN
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; Blotting, Western; Fetus; Humans; Immunoblotting; Liver; embryology; metabolism; Proteoglycans; metabolism
- From: Chinese Journal of Applied Physiology 2007;23(1):102-105
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo detect the expression of a novel protein, hemangiopoietin (HAPO), in the human fetal liver at the protein level.
METHODSMonoclonal antibodies (moAbs) against HAPO were produced by traditional hybridoma technique. Their affinities were calculated from the results of non-competitive ELISA and the antibodies were purified by protein G affinity chromatography. Expression of HAPO in the liver was detected by SDS-PAGE and Western blot.
RESULTSFive strains of moAbs were screened out in total, among which three were IgG1 and the other two were IgM. Their light trains were all belonged to kappa. The relative affinities of the three IgG1 form moAbs were 3.06 x 10(9) mol/L, 6.07 x 10(8) mol/L and 1.71 x 10(10) mol/L respectively. After purification, the purity of the moAbs could reach more than 99%. HAPO expression was detected at the protein level in the human fetal liver, and the apparent molecular weight of the nature HAPO was very close to but a little higher than our recombinant one.
CONCLUSIONHAPO was expressed in the human fetal liver at the protein level.
