Streptomycin antibodies

An indirect competitive ELISA has been also developed for the determination of streptomycin and dihydrosticptomyciri in milk (24). Prior to the analysis, the milk sample was skimmed and treated with oxalic acid. The antiserum was raised in rabbits using streptomycin linked to a bacterial protein as the antigen. To perform the test, microtiter plates were coated with streptomycin, and antiserum and milk samples were mixed to be added in the wells where they were incubated for 1 h. Depending on the amount of residues in the sample, more or less antibody remained available for binding to the streptomycin coat. A pig antirabbit antibody-enzyme conjugate was subsequently added and incubated for 90 min. Using a suitable substrate, streptomycin and dihydrostreptomycin could be detected down to 1.6 ppb, whereas quantification could be made possible up to 100 ppb when samples were used undiluted. 

Diluted antibody solutions can be reused multiple times They should be stored at 4°C after additional aliquots of penicillin/streptomycin and sodium azide have been added Some workers believe that the amount of nonspecific (background) staining on Western blots diminishes as antibody solutions are reutilized Antibody solutions are discarded or supplemented with additional antibody when the intensity of the specific signal begins to diminish 10. Choice of wash buffer after incubation with primary antibody 2M urea is included m the suggested wash buffer to diminish nonspecific binding. Alternatively, some... 

Excerpta Medica, Amsterdam London New York, pp 460-466 Girard JP, Schwartz H (1967) Serum hemagglutinating antibodies in streptomycin allergy. Med Pharmacol Exp 17 466-474... 

The 30 kDa protein is encoded by nuclear DNA, and precursor protein is transported from cytoplasm to chloroplast. As the protein is located at the lumenal surface of the thylakoid, it must also cross thylakid membrane before it is incorporated into the PS II complex properly. Identification of the precursor protein has been carried out with isolation of mRNA and identification of the translation product. Transit peptide of several kilodalton is shown to be attached to the amino terminal of the mature protein. We tried to identify the precursor protein of the 30 kDa protein with Euglena cells whose chloroplast is functionally inactivated. Treatment of the cells with streptomycin under illumination induced a protein of 36 and 52 kDa which were cross-reactable with the antibody against spinach 33 kDa protein. The proteins were also detected when chloroplast-less mutant was illuminated. The 36 kDa protein is probably a precursor of the 30 kDa protein. The 52 kDa protein was dissociated into 30 kDa protein and some other protein(s) with SDS-PAGE with 8 M urea, and it is possible that we detect a complex form of the precuresor or mature 30 kDa porotein and the protein(s) which may stabilize the free 30 kda protein or assist the 30 kDa protein in transportation across the envelope and/or thylakoid membranes. Analysis is now being carried out to elucidate the characteristics of the precursor proteins. 

Wutz et al. (2011) developed for the first time a multianalyte immunoassay based on an automated flow-through CL microarray technique for identification and quantification of antibiotic derivatives in honey samples using regenerable antigen microarrays, an indirect competitive immunoassay format using horseradish peroxidase (HRP)-labeled antibodies and CL read-out with a CCD camera. The method allows the analysis of four analytes (enrofloxacin, sulfadiazine, sulfamethazine, and streptomycin) simultaneously in 8 min with adequate recoveries and without purification or extraction. Due to the regenerability of the microarray each chip could be individually calibrated before the analysis and allowed more than 40 assays, which reduces the costs per analysis and permits an automated work flow in routine laboratories. 

Spin the suspension at 269 for 5 min, remove carefully the supernatant, and resuspend the pelleted mucosa in 1 mL DMEM/FBS plus 1 % penicillin/streptomycin. Add 8 pL anti-Thy 1.2 antibody in the solution to obtaining a 1 125 dilution. Place the 15 mL tube in a shaker at slow speed for 30 min at room temperature to allow anti-Thy 1.2 to reach specific epitopes of fibroblasts. Centrifuge the suspension at 269 x for 5 min at 4 °C and remove the supernatant. Dilute rabbit serum complement (1 mg/mL) according to manufacturer s instructions, pass the solution through a sterile syringe filter, and add 100 pL penicillin/ streptomycin. Resuspend the pellet in 1 mL of the rabbit serum complement solution described above and place again in shaking for 30 min. 

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