Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cryostat sections from rat gracilis muscles were incubated with different biotinylated lectins: Con A (Concanavilin A), WGA (Wheat germ agglutinin), SBA (soybean agglutinin), GS I and GS II (Griffonia simplicifolia agglutinin), LCA (Lens culinaris agglutinin), PNA (peanut agglutinin) and PSA (Pisum sativum agglutinin). The sections were subsequently treated with alkaline phosphatase conjugated avidin. The lectin binding sites were visualized after incubation in substrate media containing: (1) 5-bromo-4-chloro indoxyl phosphate and Nitro Blue tetrazolium or copper sulphate; (2) naphthol AS-MX phosphate or naphthol AS-BI phosphate and various types of diazonium salts; (3) alpha-naphthylphosphate and Fast Blue BB; (4) beta-glycerophosphate according to the method of Gomori. The results obtained with the alkaline phosphatase methods were compared with those seen with a streptavidin-horseradish peroxidase procedure. Several chromogen protocols for visualizing alkaline phosphatase activity showed differences in the ability to detect lectin binding sites. A sarcoplasmic reaction was evident for Con A, GS II, WGA, LCA, and PSA after incubation in the indoxyl phosphate medium. Sarcoplasmic reaction for GS II was also noticed after incubation with naphthol AS-MX Fast Blue BB and beta-glycerophosphate. The latter substrate also gave rise to a sarcoplasmic Con A reaction. With the indoxylphosphate tetrazolium salt method some muscle fibres showed a very strong intracellular reaction after incubation with Con A and GS II while the staining intensity was weak in other fibres. The same muscle fibres were stained with PAS. No sarcoplasmic reactions were observed with either naphthol phosphate media or with the diaminobenzidine peroxidase methods. Further, the staining of the muscle fibre periphery, connective tissue, an capillaries was intensified using the indoxyl method. The indoxylphosphate-tetrazolium salt method seems to be suitable for future investigations of lectin binding sites in muscle sections.
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PMID:Lectin binding in skeletal muscle. Evaluation of alkaline phosphatase conjugated avidin staining procedures. 171 10

Introduction: This study aimed at evaluating the anti-osteosarcoma activity of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) pulsed with heat shock protein 70-peptide complexes (Hsp70-PCs). Materials and methods: Human recombinant Hsp70 expression was analyzed using thin layer scanning and Western blot assay. Tumor antigens from Saos-2 cells were extracted to reconstitute Hsp70-PCs. Maturation of cord blood-derived DC was evaluated by alkaline phosphatase-anti-alkaline phosphatase kit and inverted microscope. The anti-osteosarcoma activity of CTLs evoked by DCs loaded with Hsp70-PCs was determined using Thiazolyl Blue Tetrazolium Bromide (MTT) assay. Results: Hsp70 protein level in BL21 (DE3) increased in a time-dependent manner after induction. The expression of surface markers was upregulated and a typical dendritic morphology was observed in mature DCs. Allogeneic CTLs exhibited strong cytotoxic activity against Saos-2 cells. Conclusion: Our in vitro experiment demonstrated the potent induction of cytotoxic activity against osteosarcoma using DC-based vaccine loaded with Hsp70-PCs.
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PMID:In Vitro Generation of Anti-Osteosarcoma Cytotoxic Activity Using Dendritic Cells Loaded with Heat Shock Protein 70-Peptide Complexes. 3095 40