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1) Product Images from "A Novel Immunomodulatory Mechanism Dependent on Acetylcholine Secreted by Human Bone Marrow-derived Mesenchymal Stem Cells"
Article Title: A Novel Immunomodulatory Mechanism Dependent on Acetylcholine Secreted by Human Bone Marrow-derived Mesenchymal Stem Cells
Journal: International Journal of Stem Cells
Figure Legend Snippet: The proposed model of an immunomodulatory mechanism utilized by human MSCs in an inflammatory milieu. The inflammatory conditions drive human MSCs to adopt a neuronlike phenotype. It is probable that the expression of neurotrophins such as NGF and BDNF in human AICs and the presence of NRs on MSCs are associated with these changes in MSCs. Furthermore, the inflammatory milieu probably induces the expression of nAChR including nAChR α 7, which participates in the negative regulation of activated lymphocytes. Neuronlike MSCs stimulated by neurotrophins and unknown factors secrete ACh, which binds to AICs via nAChR α 7, thereby inhibiting the proliferation and function of AICs.
Techniques Used: Expressing
Figure Legend Snippet: Inflammatory conditions induce a cholinergic-neuron–like phenotype in MSCs and nAChRs in activated PBMCs. (a) Immunofluorescence staining of ChAT, GABA, and TH in MSCs cocultured with activated PBMCs (48 h). (b) qPCR was carried out to quantify ChAT expression in MSCs after inflammatory stimulation for 24 h. (c) Western blotting confirmed ChAT expression in the whole MSC extract (20 μ g) after coculture for 48 h. (d) ACh and choline concentration was measured in the CM obtained from PBMCs alone (P or P o ), PHA-activated PBMCs (P PHA ), or MLR (P and P o ) culture without or with MSCs (n=3). (e) ACh and choline concentration was measured in the CM obtained from PBMCs alone (P or P o ) or MLR (P and P o ) culture without or with MSCs for 48 h. “MLR sup.” is the supernatant from the MLR without MSCs. “(MLR+MSC) sup.” is the supernatant from MLR with MSCs (n=3). (f) RT-PCR analysis of several nAChR subunits in activated PBMCs. (g) qPCR was carried out to assess nAChR α5 expression in activated PBMCs after incubation for 24 h. (h) qPCR was performed to measure nAChR α7 expression in activated PBMCs after incubation for 24 h. (i) An increase in the protein expression of the nAChR α 7 subunit in activated PBMCs was confirmed by western blotting of whole MSC extracts prepared after MLR or PHA stimulation for 48 h.
Techniques Used: Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Concentration Assay, Reverse Transcription Polymerase Chain Reaction, Incubation