Preparation and application of rabbit polyclonal antibody against human lactate dehydrogenase C4(LDHC4).
- Author:
Axiu HUANG
1
,
2
;
Caiyun DENG
3
;
Siyu YANG
3
;
Qinglian ZHANG
4
Author Information
1. School of Laboratory Medicine, Chengdu Medical College, Chengdu 610500
2. Medical laboratory, The Second People's Hospital of Jiangyou, Mianyan 621050, China.
3. School of Laboratory Medicine, Chengdu Medical College, Chengdu 610500, China.
4. School of Laboratory Medicine, Chengdu Medical College, Chengdu 610500, China. *Corresponding author, E-mail: qlzhang80@163.com.
- Publication Type:Journal Article
- MeSH:
Humans;
Rabbits;
Animals;
Escherichia coli/genetics*;
Antibodies;
Enzyme-Linked Immunosorbent Assay;
L-Lactate Dehydrogenase/metabolism*;
Blotting, Western;
Antibody Specificity
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(12):1118-1124
- CountryChina
- Language:Chinese
-
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 1:51 200. The antibody can be used for Western blot and immunohistochemistry. Conclusion The specific rabbit anti-human LDHC4 polyclonal antibody is successfully prepared.
