Eaat2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Small Extracellular Vesicles in Rat Serum Contain Astrocyte-Derived Protein Biomarkers of Repetitive Stress"
Article Title: Small Extracellular Vesicles in Rat Serum Contain Astrocyte-Derived Protein Biomarkers of Repetitive Stress
Journal: International Journal of Neuropsychopharmacology
Figure Legend Snippet: Aldolase C tagged with green fluorescent protein (C-GFP) expressed in forebrain astrocytes is detected in extracellular vesicles (EVs) isolated from the blood. (a) Scheme of the in utero electroporation. Forebrain astrocytes were transduced by in utero electroporation with aldolase C-GFP or GFP. Plasmids were injected into the left lateral ventricle on embryonic day 18.5. The orientation of the electrodes used to apply the voltage pulse is shown. (b) Immunohistofluorescent detection of glial fibrillary acid protein (GFAP) and GFP in coronal brain slices indicated that cells positive for both proteins were detected in the borders of the lateral ventricles (LV, indicated by arrows), and (c) in the hilus of the dentate gyrus (DG). Gr, granule cells. (d) Aldolase C (left) or GFP (right) was detected in astrocyte homogenates (AH) electroporated with aldolase C-GFP or in sEVs isolated from the serum of these animals. Note that in the sEVs, the modified form of aldolase C was detected (~ 55 kDa), as well as the recombinant protein (~ 70 kDa), which was also visible with the GFP antibody. (e) Aldolase C (left) or GFP (right) was detected in the astrocyte homogenates (AH) that were electroporated with GFP or in extracellular vesicles (sEVs) isolated from the serum of these animals. Note that in sEVs, the modified form of aldolase C was detected (~ 55 kDa), while in these animals, no GFP could be detected in the EVs. Observations were conducted in n = 5 independent animal groups (n = 4 rats per group for blood collection). (f) EVs bearing the glial glutamate transporter EAAT2 in their membrane are enriched in aldolase C. EVs were immunoisolated in nondenaturating conditions with an antibody directed against an extracellular epitope of EAAT2. The input lane was loaded with 8% of the EVs used for the precipitation procedure (i.e., 20 µg). Note that EAAT2 can only be detected in the input after overexposure of the same blot (right lanes), revealing a huge enrichment of EAAT2-bearing vesicles in the isolated material that in turn also contains higher aldolase C levels compared with the input EVs.
Techniques Used: Isolation, In Utero, Electroporation, Injection, Modification, Recombinant