Analysis conditions for proteomic profiling of mammalian tissue and cell extracts with antibody microarrays.
Antibody microarrays are a developing tool for global proteomic profiling. A protocol was established that permits robust analyses of protein extracts from mammalian tissues and cells rather than body fluids. The factors optimized were buffer composition for surface blocking, blocking duration, protein handling and processing, labeling parameters like type of dye, molar ratio of label versus protein, and dye removal, as well as incubation parameters such as duration, temperature, buffer, and sample agitation.
Alhamdani
Mohamed Saiel Saeed
MS
Division of Functional Genome Analysis, Deutsches Krebsforschungszentrum (DKFZ), Heidelberg, Germany. m.alhamdani@dkfz.de
Schröder
Christoph
C
Hoheisel
Jörg D
JD
D000906:Antibodies / Q000737:chemistry*; D002021:Buffers; D002457:Cell Extracts / Q000737:chemistry; D005456:Fluorescent Dyes; D006801:Humans; D007202:Indicators and Reagents; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D046888:Tissue Array Analysis / Q000379:methods*
D000906:Antibodies; D002021:Buffers; D002457:Cell Extracts; D005456:Fluorescent Dyes; D007202:Indicators and Reagents