Preparation and application of rabbit polyclonal antibody against human lactate dehydrogenase C4(LDHC4) / 细胞与分子免疫学杂志
Chinese Journal of Cellular and Molecular Immunology
;
(12): 1118-1124, 2023.
Artículo
en Chino
| WPRIM
| ID: wpr-1009463
ABSTRACT
Objective To prepare rabbit polyclonal antibody specifically against human lactate dehydrogenase C4 (LDHC4). Methods Site-directed mutation was performed by PCR to generate the mutated LDHC gene, and the mutated gene was ligated into the pET-28a vector to form the pET-28a-LDHC recombinant expression vector. The recombinant vector was introduced into E. coli BL21 (DE3), and LDHC4 protein was obtained by induced expression. The recombinant protein was used as an antigen to immunize New Zealand rabbits, and the antiserum was obtained after three boosted immunizations. The titer of the antiserum against LDHC4 were detected by ELISA. Western blot was used to detect the specificity of the antiserum, and immunohistochemistry was used to detect the expression of LDHC4 in human triple-negative breast cancer tissue. Results A specific rabbit anti-human LDHC4 polyclonal antibody was obtained with an antibody titer of 151 200. The antibody can be used for Western blot and immunohistochemistry. Conclusion The specific rabbit anti-human LDHC4 polyclonal antibody is successfully prepared.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Ensayo de Inmunoadsorción Enzimática
/
Western Blotting
/
Escherichia coli
/
L-Lactato Deshidrogenasa
/
Anticuerpos
/
Especificidad de Anticuerpos
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Cellular and Molecular Immunology
Año:
2023
Tipo del documento:
Artículo
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