Purification and characterization of a highly specific polyclonal antibody against human extracellular signal-regulated kinase 8 and its detection in lung cancer.
Na-Li CaiAndy T Y LauFei-Yuan YuDan-Dan WuLi-Juan DaiHai-Ying MoChang-Min LinYan-Ming XuPublished in: PloS one (2017)
Extracellular signal-regulated kinase 8 (ERK8), proposed as a novel potential therapeutic target for cancer, has been implicated in cell transformation, apoptosis, the protection of genomic integrity, and autophagy. To facilitate ERK8 research, a highly specific anti-ERK8 antibody is needed. In this article, we use the Immune Epitope Database and Analysis Resource online tool to predict B-cell epitopes of human ERK8 protein, and choose a 28 aa-peptide sequence to generate the GST-ERK8(28aa) fusion protein as the antigen for developing polyclonal antibody against ERK8. The specificity and sensitivity of anti-ERK8 antibody were robustly validated by immunoblotting, immunocytochemical and immunohistochemical analyses; and we found that both the endogenous and ectopically-expressed human ERK8 proteins can be recognized by our anti-ERK8 antibody. This suggested that our characterized anti-ERK8 antibody will be a valuable tool for the elucidation of the distribution of ERK8 at cellular and histological levels. Finally, our tissue array analysis also demonstrated that the ERK8 protein was localized in both the nucleus and cytoplasm of human lung cancers.
Keyphrases
- signaling pathway
- pi k akt
- cell proliferation
- endothelial cells
- cell cycle arrest
- oxidative stress
- healthcare
- cell death
- social media
- risk assessment
- emergency department
- endoplasmic reticulum stress
- single cell
- stem cells
- induced pluripotent stem cells
- mesenchymal stem cells
- high resolution
- gene expression
- bone marrow
- health information
- copy number
- lymph node metastasis
- drug induced
- data analysis
- squamous cell