Cell-Free Bead-Based Detection of Total and Phosphorylated Proteins in Plasma and Cell Lysates: Detection of FLT3
Frequently direct measurement of proteins or their phosphorylation in intact cells is not possible, for instance, when cells are too few, frozen, or subject to degradation. We have demonstrated that tumor cells pour their DNA, RNA, and protein content into circulation because of turnover and breakdown of cell structures. Proteins in solution most likely circulate as complexes, which protects them from degradation. We describe a cell-free, bead-based method that takes advantage of this phenomenon. Our approach is based on immunoprecipitation of the protein of interest on the surface of beads, followed by detection of the protein or its modification (phosphorylation) using a secondary antibody labeled with phycoerythrin at a 1∶1 ratio. Fms-like tyrosine kinase-3, which is mutated in majority of cases of acute myeloid leukemia, is used as an example. This method could be applied to the quantitation of several other proteins without the need for intact cells.
- 免疫金银染色法(immunogold-silver taining,IGSS)
- 中性粒细胞碱性磷酸酶活性组织化学检查法
- 人前列腺特异性抗原(PSA)酶联免疫分析(ELISA)
- 细胞因子的作用特点
- 抗血清的制备
- Transwell Chemotaxis
- Recombinant Fusion Toxins Directed Against the Human Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) Receptor
- Production of Polyclonal Antibodies in Rabbits
- Evaluation of Viral Interference with MHC Class I-Restricted Antigen Processing and Presentation Using a Flow Cytometry-Based Ap
- CD66a/CD66b/CD66c/CD66d/CD66e/CD66f/CD67分子