ABSTRACT
The transforming growth factor-beta (TGF-beta) cytokine family has important and complex effects on many biologic processes. Mammals have three TGF-beta isoforms which differ in their primary amino acid sequence, receptor binding characteristics, distribution, and function. Characterization of TGF-beta production and localization is critically dependent upon appropriate reagents, including antibodies. We have analyzed the isoform specificity of eight commercially-available TGF-beta antibodies, including one monoclonal antibody and seven polyclonal antibodies. We carried out semi-quantitative Western blot analysis using recombinant TGF-beta1, beta2, and beta3 as targets. We found that sensitivity and isoform specificity are dependent in part upon the presence or absence of reducing conditions. The antibodies tested showed a broad range of sensitivity, with an ability to detect 50 pg to 20 ng. Cross-reactivity with another, incorrect isoform was seen with several antibodies, and ranged from 0.2% to 42%. Nevertheless, we identified TGF-beta antibodies directed against each isoform which provide moderate-to-high sensitivity and specificity when used in Western blot analysis. These results may have relevance for investigators who wish to detect particular TGF-beta isoforms with techniques other than Western blot analysis, particularly when these techniques involve denatured proteins.
Subject(s)
Protein Isoforms/immunology , Transforming Growth Factor beta/immunology , Animals , Antibodies/chemistry , Antibody Specificity , Antigens/immunology , Blotting, Western , Cross Reactions , HumansABSTRACT
The 60 year old man was admitted because of aphasia and hemiparesis. After cranial computed tomography 15 ml parietal hematoma was removed by stereotaxic biopsy. The patient had hyperpyrexia, combined mitral vitium and atrial fibrillation. There was no symptom of gastroenteritis. Salmonella enteritidis was cultured from blood three times. The vegetation was proved by transoesophageal echocardiography. Ampicillin + gentamycin, amoxicillin-clavulanate + amikacin therapy was ineffective, respectively. During ciprofloxacin therapy of usual dose ceased the toxicosis and hyperpyrexia, but remained fever to 38.5 degrees C. During 750 mg ciprofloxacin t. i. d. intravenous followed 750 mg t. i. d. per os plus 1.5 g cefuroxin t. i. d. intravenous for 46 days became the patient afebrile and the vegetation was disappeared. No side effect was observed with ciprofloxacin of unusual high daily dose.