Fluorescent Protein Specific Nanotraps to Study ProteinProtein Interactions and Histone-Tail Peptide Binding
Fluorescent proteins are widely used to study protein localization and protein dynamics in living cells. Additional information on peptide binding, DNA binding, enzymatic activity, and complex formation can be obtained with various methods including chromatin immunoprecipitation (ChIP) and affinity purification. Here we describe two specific GFP- and RFP binding proteins based on antibody fragments derived from llama single domain antibodies. The binding proteins can be produced in bacteria and coupled to monovalent matrixes generating so-called Nanotraps. Both Nanotraps allow a fast and efficient (one-step) isolation of fluorescent fusion proteins and their interacting factors for biochemical analyses including mass spectroscopy and enzyme activity measurements. Here we provide protocols for precipitation of fluorescent fusion proteins from crude cell extracts to identify and map protein–protein interactions as well as specific histone tail peptide binding in an easy and reliable manner.
- 血小板计数法(直接试管法)
- HLA等位基因序列特异性探针(SSO)及其标记
- 干燥症
- 免疫应答(三):免疫耐受-- 免疫耐受的发现
- 细胞因子的临床意义
- 酶连接的免疫吸收剂化验(ELISA, Enzyme Linked ImmunoSorbent Assay)
- Mast Cell Apoptosis
- ELISA (Enzyme-Linked ImmunoadSorbent Assay)
- Determination of Antigen and Fine Peptide Specificity of EBV-Specific CTLs
- Detection of High-Risk Mucosal Human Papillomavirus DNA in Human Specimens by a Novel and Sensitive Multiplex PCR Method Combine