Generation of a rat monoclonal antibody specific for CHD2.
CHD2 is a member of the CHD family that contains chromodomain, helicase domain as well as DNA-binding domain. The CHD family is involved in gene expression and transcription by ATP-dependent chromatin remodeling. Analysis of mutant mouse revealed that CHD2 is involved in development as well as hematopoiesis, which suggests the involvement of CHD2 in gene expression. However, CHD2 has not yet been analyzed biochemically as there is no specific antibody against it. Here, we report on the establishment of specific monoclonal antibody (MAb) against CHD2 utilizing a rat medial iliac lymph node method. Through cell immunostaining utilizing established MAb to CHD2, we confirmed that CHD2 was localized in euchromatin. Additionally, IP-Western revealed that the expression level of full-length CHD2 did not change during the differentiation stage. Additionally, a specific signal was confirmed around 95 kDa at the undifferentiated stage. This clearly indicated that CHD2 was involved in specific gene expression at this stage. Thus, this antibody can contribute to elucidating the function of CHD2 in cell expression.
Harada
Akihito
A
Department of Epigenetics, SSP Stem Cell Unit, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Yoshimura
Saori
S
Odawara
Jun
J
Azuma
Masayuki
M
Okada
Seiji
S
Nakamura
Mako
M
Tachibana
Taro
T
Ohkawa
Yasuyuki
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005786:Gene Expression Regulation / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology*; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D051381:Rats
D000911:Antibodies, Monoclonal; C514205:Chd2 protein, mouse; D004268:DNA-Binding Proteins