Generation of monoclonal antibodies against recombinant AtSIZ1.
Post-translational modifications of target proteins by small ubiquitin-like modifier (SUMO) proteins modulate many cellular processes in yeast and animals. Here we present the development of monoclonal antibodies (MAb) and polyclonal antibodies (PAb) against Arabidopsis SIZ1 (AtSIZ1) protein with high specificity. Mice were immunized with recombinant AtSIZ1 protein for generating monoclonal antibodies via the classic hybridoma production technique. Anti-AtSIZ1 MAb and PAb were able to detect endogenous AtSIZ1 in Arabidopsis wild type and its complementary line formed by transforming C-siz1-2 mutant with construct containing the AtSIZ1 gene under the control of the native promoter, but not the siz1-2 deletion mutant. These results show that these anti-AtSIZ MAbs are highly sensitive to detect endogenous AtSIZ1 and can be used for immunoblotting and other experimental methods. The new anti-AtSIZ1 MAbs will be essential tools used to investigate the role of AtSIZ1 in plant developmental biology.
Kim
Kangchang
K
Division of Applied Life Science (Brain Korea 21 Program), PMBBRC, EB-NCRC, Gyeongsang National University, Jinju City, Korea.
Bae
Suyoung
S
Hong
Jaewoo
J
Choi
Jida
J
Ryoo
Soyoon
S
Jhun
Hyunjhung
H
Lee
Siyoung
S
Her
Erk
E
Hong
Kwangwon
K
Kim
Soohyun
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification* / Q000627:therapeutic use; D000918:Antibody Specificity; D017360:Arabidopsis / Q000235:genetics / Q000276:immunology*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology*; D008025:Ligases / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D029681:Arabidopsis Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D008025:Ligases; C502552:SIZ1 protein, Arabidopsis