[Extraction and purification of porcine amelogenin and preparation for the polyclonal amelogenin antibody].
OBJECTIVE
To prepare the polyclonal antibody to amelogenin.


METHODS
The fetal porcine dental enamel was collected. Enamel matrix protein was extracted in 4M guanidine HCl (pH 7.4) with protease inhibitors present. Polyacrylamide gel filtration was included to isolate amelogenin from the initial dissociated extraction. The purified amelogenin conjugated with or without complete/incomplete Freund's adjuvant was then used to immunize the rabbits subcutaneously or intravenously. The specific IgG antibody was further purified by DE-52 cellulose. The working concentration of IgG antibody was determined through ELISA test.


RESULTS
The Gel filtration showed that amelogenin components is at molecular weights of 15 kD and 13 kD apparently, which was consistent with those described before. The ELISA results showed that the working concentration for IgG was 1:1000.


CONCLUSIONS
The antibody prepared in this study can be used for the detection of amelogenin.
Xu
Chenrong
C
Department of Oral Medicine, West China College of Stomatology, Sichuan University, Chengdu 610041, China.
Yang
Ailing
A
Zhang
Jincai
J
D053523:Amelogenin; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D003743:Dental Enamel / Q000737:chemistry*; D003746:Dental Enamel Proteins / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis; D011817:Rabbits; D013552:Swine; D014083:Tooth Germ / Q000737:chemistry
D053523:Amelogenin; D000911:Antibodies, Monoclonal; D003746:Dental Enamel Proteins; D007074:Immunoglobulin G