THE PRODUCTION AND CHARACTERIZATION OF MURINE MONOCLONAL-ANTIBODIES TO HUMAN GADD45 RAISED AGAINST A RECOMBINANT PROTEIN

被引:5
作者
KILPATRICK, KE
CARRIER, F
SMITH, ML
CHEN, CY
LEE, AJ
RUSNAK, DW
KASTAN, MB
FORNACE, AJ
CHAMPION, BR
GILMER, TM
SU, JL
机构
[1] NCI,DCT,DTP,MOLEC PHARMACOL LAB,BETHESDA,MD 20892
[2] JOHNS HOPKINS UNIV,CTR ONCOL,BALTIMORE,MD 21287
[3] GLAXO INC,RES INST,DEPT CELL BIOL,RES TRIANGLE PK,NC 27709
来源
HYBRIDOMA | 1995年 / 14卷 / 04期
关键词
D O I
10.1089/hyb.1995.14.355
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The production of two different murine monoclonal antibodies to human Gadd45, a protein that is induced in response to DNA damage, is reported, Antibodies were generated in a SJL mouse using a recombinant form of the human Gadd45 protein. Monoclonal antibody 4TCYA1, which recognizes the denatured form of human Gadd45 in Western blots, was selected based upon the recognition of Gadd45 induced by functional p53 in the human myeloid leukemia cell line, ML-1, A second monoclonal antibody, designated 30T,14, immunoprecipitates native human Gadd45 in lysates produced from RKO cells, a colorectal carcinoma cell line that expresses relatively high basal levels of Gadd45, as well as from cell lysates made from ML-1 cells after exposure to ionizing irradiation (IR), Since 4TCYA1 fails to immunoprecipitate Gadd45, and 30T,14 fails to bind to IR-induced Gadd45 in immunoblotting, these two monoclonal antibodies probably recognize different epitopes.
引用
收藏
页码:355 / 359
页数:5
相关论文
共 18 条
[1]   WILD-TYPE BUT NOT MUTANT P53 IMMUNOPURIFIED PROTEINS BIND TO SEQUENCES ADJACENT TO THE SV40 ORIGIN OF REPLICATION [J].
BARGONETTI, J ;
FRIEDMAN, PN ;
KERN, SE ;
VOGELSTEIN, B ;
PRIVES, C .
CELL, 1991, 65 (06) :1083-1091
[2]   ELECTROPHORETIC TRANSFER OF PROTEINS AND NUCLEIC-ACIDS FROM SLAB GELS TO DIAZOBENZYLOXYMETHYL CELLULOSE OR NITROCELLULOSE SHEETS [J].
BITTNER, M ;
KUPFERER, P ;
MORRIS, CF .
ANALYTICAL BIOCHEMISTRY, 1980, 102 (02) :459-471
[3]  
BURNETTE WN, 1981, ANAL BIOCHEM, V112, P195, DOI 10.1016/0003-2697(81)90281-5
[4]  
CANMAN CE, 1994, CANCER RES, V54, P5094
[5]  
CARRIER F, 1994, J BIOL CHEM, V269, P32672
[6]   DNA DAMAGE-INDUCIBLE TRANSCRIPTS IN MAMMALIAN-CELLS [J].
FORNACE, AJ ;
ALAMO, I ;
HOLLANDER, MC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (23) :8800-8804
[7]   MAMMALIAN GENES COORDINATELY REGULATED BY GROWTH ARREST SIGNALS AND DNA-DAMAGING AGENTS [J].
FORNACE, AJ ;
NEBERT, DW ;
HOLLANDER, MC ;
LUETHY, JD ;
PAPATHANASIOU, M ;
FARGNOLI, J ;
HOLBROOK, NJ .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (10) :4196-4203
[8]   A MAMMALIAN-CELL CYCLE CHECKPOINT PATHWAY UTILIZING P53 AND GADD45 IS DEFECTIVE IN ATAXIA-TELANGIECTASIA [J].
KASTAN, MB ;
ZHAN, QM ;
ELDEIRY, WS ;
CARRIER, F ;
JACKS, T ;
WALSH, WV ;
PLUNKETT, BS ;
VOGELSTEIN, B ;
FORNACE, AJ .
CELL, 1992, 71 (04) :587-597
[9]  
KEARNEY JF, 1979, J IMMUNOL, V123, P1578
[10]  
KUERBITZ SJ, 1992, P NATL ACAD SCI USA, V16, P7491