Steinitz M, Tamir S
Department of Pathology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
J Immunol Methods. 1995 Nov 16;187(1):171-7. doi: 10.1016/0022-1759(95)00181-9.
A method is described for producing 1-3 microns sized particles of nitrocellulose (NC) which are able to absorb protein. Protein is absorbed onto preformed particles made by first dissolving a sheet of nitrocellulose paper in DMSO, and then precipitating it with sodium carbonate/bicarbonate buffer. The efficiency of binding is the same as that of an equivalent sheet of non-processed NC filter paper. Antibodies absorbed onto preformed particles are not exposed to DMSO and carbonate buffer and therefore retain a high antigen binding capacity. Antigen and antibody-absorbed NC preformed particles were used to capture antibody and antigen, respectively. Using lysis buffer, the captured antibodies and antigens were readily released from the NC particles. This makes the latter an appropriate matrix for immunoprecipitation assays either for an antigen or for specific antibody. Antigen-coated NC particles were specifically aggregated ('agglutinated') by specific antibodies and thus can be used in semi-quantitative tests.
描述了一种制备能够吸收蛋白质的1 - 3微米大小硝化纤维素（NC）颗粒的方法。蛋白质被吸收到预先形成的颗粒上，这些颗粒是通过首先将一片硝化纤维素纸溶解在二甲基亚砜（DMSO）中，然后用碳酸钠/碳酸氢钠缓冲液沉淀而制成的。结合效率与同等的未处理NC滤纸相同。吸收到预先形成颗粒上的抗体未暴露于DMSO和碳酸盐缓冲液中，因此保留了高抗原结合能力。抗原和抗体吸收的NC预先形成颗粒分别用于捕获抗体和抗原。使用裂解缓冲液，捕获的抗体和抗原很容易从NC颗粒中释放出来。这使得后者成为用于抗原或特异性抗体免疫沉淀测定的合适基质。抗原包被的NC颗粒被特异性抗体特异性聚集（“凝集”），因此可用于半定量测试。
