The development of ELISA is based on the immobilization and enzyme labeling of antigen or antibody. The immunological activity of antigen or antibody remains after the immobilization. And enzyme-labeled antigen or antibody remains both their immunological activity and enzymatic activity. Materials: Antigen Serum Reagent and kit: Carbonate coating buffer PBST BSA Equipment and Supplies: 96-Well ELISA plates 4℃refrigerator Incubator Spectrophotometer Procedure: (1) Coating antigen 1. Dilute the antigen into 10-20μg/ml with 50 mM carbonate coating buffer (pH9.6)…
Protocol
Principle and Protocol of Chromatin Immunoprecipitation (ChIP)
Introduction: 1. Applications of ChIP: (1) Histone modifying enzyme antibody as “biomarkers”; (2) Analysis of transcriptional regulation; (3) Drug development research; (4) Analysis of DNA damage and apoptosis. 2. Details: The eukaryotic genomic DNA exists in the form of chromatin. Therefore, the study of interactions between protein and DNA in chromatin environment is the basic way to clarify the mechanism of gene expression in eukaryotes. Chromatin immunoprecipitation (ChIP) is currently…
Principle and Protocol of Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE)
Principle The concentration of polyacrylamide gels can be prepared as required in two electrophoresis systems —called “continuous system” and “discontinuous system”. The biggest feature of “discontinuous system” lies in its greatly improved sample separation resolution. Main features of this electrophoresis are: (1) Use of two gel systems with different concentrations; (2) Solution composition and pH are different for the preparation of the two gel and are also different from electrophoresis…