[Preparation of a polyclonal antibody against N-terminal of 1A6/DRIM with MAPs method].
OBJECTIVE
To obtain the antibody against N-terminal of 1A6/DRIM, and thereafter get the profile of 1A6/DRIM expression in different cell lines.


METHODS
The N-terminal of 1A6/DRIM (aa 577-714) was cloned into pGEX-4T-3. Multiple antigenic peptides   MAPs   (aa638-661) was synthesized as the antigen with Fmoc/PyBOP method. Rabbits were immunized by injecting the MAPs and the immunized sera were analyzed with ELISA and Western Blot. The Western Blot and immunofluorescence were performed to analyze the expressing profiles of the 1A6/DRIM in different tumor cell lines.


RESULTS
The antibody specifically recognized the full length of 1A6/DRIM as a 310 kDa band, which was also recognized by C-terminal monoclonal antibody shown by Western Blot. 1A6/DRIM is expressed in multiple tumor cell lines and mainly located in the nuclei.


CONCLUSIONS
Preparation of the antibody with MAPs is a useful technique when the fusion proteins can not be induced in E. coli. The antibody we got via MAPs has supplied a good tool for further studies on the functions of the novel gene 1A6/DRIM.
Department of Oncology, Beijing Hospital, Beijing 100039, China.
Ying-ying
YY
Huang
Xiao-nan
XN
Wu
Xiao-juan
XJ
Du
Tao
T
Ning
Ning
N
Li
Jun
J
Zhu
Yang
Y
Ke
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D015153:Blotting, Western; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D010455:Peptides / Q000276:immunology*; D012261:Ribonucleoproteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D036341:Intercellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D010455:Peptides; D012261:Ribonucleoproteins; C113527:UTP20 protein, human; C089108:multiple antigen peptide, Leu(7)-