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Polyclonal Serum (polyclonal + serum)
Selected AbstractsEXPRESSION OF A BEE-VENOM PHOSPHOLIPASE A2 FROM APIS CERANA CERANA IN ESCHERICHIA COLIINSECT SCIENCE, Issue 1 2004Li-rong Shen Abstract, The venomous phospholipase A2 (AcPLA2) coding reading region of the Chinese honeybee (Apis cerana cerana), which is composed of 405 bp encoding a mature glycosylated peptide with 134 amino residues was transformed into the expression vector pETblue-1. Then the recombinant vector was introduced into Escherichia coli Tuner (DE3) plac I for expression. Analysis result of SDS-PAGE showed that the expression products had a protein band of about 15kD. Detection of western blot using ant-European honeybee (Apis mellifera) phospholipase A2 (AmPLA2) polyclonal serum as the first antibody showed that the expression products appeared a special blot same as the native AmPLA2. The result demonstrated that the AcPLA2 peptide had been expressed in E. coli and the AcPLA2 has the similar antigenicity as the AmPLA2. [source] Effect of substrate size on immunoinhibition of amylase activity,JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2001Ilka Warshawsky Abstract Immunoinhibition assays are hypothesized to work by antibodies blocking substrate access to enzyme active sites. To test this hypothesis, the inhibition of amylase isoenzymes by monoclonal and polyclonal antisera was assessed using substrates of varying sizes: chromogenic sustrates 3, 5, or 7 glucose units in length, novel synthetic macromolecular substrates, and starch. The synthetic macromolecular substrates consisted of small oligosaccharide substrates linked to an inert polymer that conferred a large size to substrate molecules as determined by gel filtration chromatography. When substrate size increased, amylase activity could be inhibited equivalently by antibody concentrations that are 10‐fold lower. Progressively less polyclonal serum was required to inhibit amylase activity as substrate length increased from 3 to 5 to 7 glucose units and as size was increased by linkage to a polymer. Different effects of substrate size were observed with two monoclonal antibodies. One monoclonal antibody blocked amylase activity independent of substrate size, while another monoclonal antibody had little inhibitory effect except using starch as substrate. We conclude that use of larger substrates can expand the repertoire of inhibitory epitopes on enzymes and convert a noninhibitory antibody into an inhibitory one. J. Clin. Lab. Anal. 15:64–70, 2001. [source] Preparation of nitrocellulose (NC) immuno-affinity membrane for purification of rAPC antibodyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 6-7 2008Haixiang Sun Abstract In this study, recombinant allophycocyanin (rAPC) with a purity of 98% was transferred from a gel to a nitrocellulose (NC) membrane to develop a simple and efficient immuno-affinity membrane. Atomic force microscopy (AFM) was used to investigate the surface topography of the affinity membrane and its characterization indicated that rAPC easily forms trimers or hexamers on the membrane surface on use of the given transfer method. The hydrodynamic radius (Rh) of the rAPC aggregation was equal to 103 nm or 365 nm according to dynamic light scattering (DLS), which was in agreement with the result obtained by AFM. Based on the specific immunological reaction of antigen and antibody, anti-APC antibodies were purified from rabbit polyclonal serum in a single step. The amount of absorbed antibody was 5.79 mg/g membrane according to analysis by ELISA methods. The purity of antibodies was up to 98% according to SDS-PAGE. The adsorption-desorption cycle of rAPC was repeated six times using the same immuno-affinity membrane, and there was no significant loss in adsorption capacity. The method provides a novel and efficient immunological affinity membrane for the purification of antibodies. [source] Molecular recognition specificity of Bacillus globigii spore antibodiesLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2000P. Longchamp Western blotting methods have been used to assess the specificity of polyclonal antibodies raised against Bacillus globigii spore and vegetative cell preparations. None of the antibodies studied were completely species-specific in their recognition of spore surface epitopes. One polyclonal serum recognized several spore surface epitopes and demonstrated limited cross-reaction with the spore surface of the near-neighbour species B. subtilis. A second polyclonal serum, raised against aged spore antigens, recognized damaged spore epitopes primarily. Both of these antibodies also cross-reacted with vegetative cell epitopes present in all four Bacillus species (B. globigii, B. subtilis, B. cereus and B. anthracis) studied. [source] | ||||||||||