eaat2  (Alomone Labs)


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    Structured Review

    Alomone Labs eaat2
    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 <t>EAAT2</t> 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.
    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
    https://www.bioz.com/result/eaat2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    eaat2 - by Bioz Stars, 2022-12
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    Images

    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

    doi: 10.1093/ijnp/pyy098

    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.
    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

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    Alomone Labs anti glt1
    Anti Glt1, 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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    Alomone Labs glt1
    Adeno-associated virus (AAV)-mediated deletion of Per2 in glial cells of adult mice leads to a depression resistant behavior. (A) Fluorescent imaging of whole brains 3 weeks after no injection (left), intravenous (i.v.) injection of the engineered AAV-PHP.eB, which can pass the blood-brain barrier (BBB), containing the general CAG driver (second from left) or the glial Gfap driver (middle). The second to last brain is from an animal with i.v. injected AAV-9, which does not pass the BBB, containing the general CAG driver. The last brain (right) is from an animal injected intraperitoneally (i.p.) with the AAV-PHP.eB Gfap -driven construct. Note that only the brains of animals that received the AAV-PHP.eb i.v. display significant fluorescent signal after 3 weeks (orange and yellow color). (B) Fluorescent imaging of whole brains 2 months after injection of the AAV-PHP.eB. Note that the fluorescence is still maintained after 2 months post injection and that even the i.p. injected AAV-PHP.eB Gfap is showing signal in the brain now. (C) Sorting of neurons and astrocytes by flow cytometry from brain tissue including the nucleus accumbens (NAc). The left panel shows the removal of debris from a single cell suspension, showing the distribution of debris in the forward as well as in the side scatter (FSC and SSC, respectively). The middle panel shows the removal of CD11b + cells (microglia) from the cell suspension (lower left corner from left panel). The CD11b - cells (bottom half from middle panel) were then sorted into two distinct cell populations corresponding to astrocytes <t>(GLT1</t> + /CD90.2 - ) and neurons (CD90.2 + /GLT1 - ) (right panel). (D) PCR analysis of astrocytes and neurons from the cell sorting. Microglia (CD11 + ) as well as astrocytes (GLT1 + /CD90.2 - ), but not neurons (CD90.2 + /GLT1 - ) from PHP.eB Gfap-iCre infected animals show the presence of iCre , indicating that only glia and not neurons could express iCre in order to delete Per2 in the Per2 fl/fl mice. (E) Immunohistochemistry of vG Per2 brain tissue from nucleus accumbens (NAc) isolated at ZT6. The signal for PER2 (green) mainly overlaps with neuronal NeuN signal (red) giving rise to the yellow color. Scale bar: 100 µm. (F) Immobility time in the forced swim test (FST) of vG Per2 (PHP.eB Gfap-iCre , green) and control (PHP.eB control, blue) animals are shown (n = 5, two-tailed t-test, *p
    Glt1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Adeno-associated virus (AAV)-mediated deletion of Per2 in glial cells of adult mice leads to a depression resistant behavior. (A) Fluorescent imaging of whole brains 3 weeks after no injection (left), intravenous (i.v.) injection of the engineered AAV-PHP.eB, which can pass the blood-brain barrier (BBB), containing the general CAG driver (second from left) or the glial Gfap driver (middle). The second to last brain is from an animal with i.v. injected AAV-9, which does not pass the BBB, containing the general CAG driver. The last brain (right) is from an animal injected intraperitoneally (i.p.) with the AAV-PHP.eB Gfap -driven construct. Note that only the brains of animals that received the AAV-PHP.eb i.v. display significant fluorescent signal after 3 weeks (orange and yellow color). (B) Fluorescent imaging of whole brains 2 months after injection of the AAV-PHP.eB. Note that the fluorescence is still maintained after 2 months post injection and that even the i.p. injected AAV-PHP.eB Gfap is showing signal in the brain now. (C) Sorting of neurons and astrocytes by flow cytometry from brain tissue including the nucleus accumbens (NAc). The left panel shows the removal of debris from a single cell suspension, showing the distribution of debris in the forward as well as in the side scatter (FSC and SSC, respectively). The middle panel shows the removal of CD11b + cells (microglia) from the cell suspension (lower left corner from left panel). The CD11b - cells (bottom half from middle panel) were then sorted into two distinct cell populations corresponding to astrocytes (GLT1 + /CD90.2 - ) and neurons (CD90.2 + /GLT1 - ) (right panel). (D) PCR analysis of astrocytes and neurons from the cell sorting. Microglia (CD11 + ) as well as astrocytes (GLT1 + /CD90.2 - ), but not neurons (CD90.2 + /GLT1 - ) from PHP.eB Gfap-iCre infected animals show the presence of iCre , indicating that only glia and not neurons could express iCre in order to delete Per2 in the Per2 fl/fl mice. (E) Immunohistochemistry of vG Per2 brain tissue from nucleus accumbens (NAc) isolated at ZT6. The signal for PER2 (green) mainly overlaps with neuronal NeuN signal (red) giving rise to the yellow color. Scale bar: 100 µm. (F) Immobility time in the forced swim test (FST) of vG Per2 (PHP.eB Gfap-iCre , green) and control (PHP.eB control, blue) animals are shown (n = 5, two-tailed t-test, *p

    Journal: bioRxiv

    Article Title: Deletion of the Clock Gene Period2 (Per2) in Glial Cells Alters Mood-Related Behavior in Mice

    doi: 10.1101/2020.12.09.417162

    Figure Lengend Snippet: Adeno-associated virus (AAV)-mediated deletion of Per2 in glial cells of adult mice leads to a depression resistant behavior. (A) Fluorescent imaging of whole brains 3 weeks after no injection (left), intravenous (i.v.) injection of the engineered AAV-PHP.eB, which can pass the blood-brain barrier (BBB), containing the general CAG driver (second from left) or the glial Gfap driver (middle). The second to last brain is from an animal with i.v. injected AAV-9, which does not pass the BBB, containing the general CAG driver. The last brain (right) is from an animal injected intraperitoneally (i.p.) with the AAV-PHP.eB Gfap -driven construct. Note that only the brains of animals that received the AAV-PHP.eb i.v. display significant fluorescent signal after 3 weeks (orange and yellow color). (B) Fluorescent imaging of whole brains 2 months after injection of the AAV-PHP.eB. Note that the fluorescence is still maintained after 2 months post injection and that even the i.p. injected AAV-PHP.eB Gfap is showing signal in the brain now. (C) Sorting of neurons and astrocytes by flow cytometry from brain tissue including the nucleus accumbens (NAc). The left panel shows the removal of debris from a single cell suspension, showing the distribution of debris in the forward as well as in the side scatter (FSC and SSC, respectively). The middle panel shows the removal of CD11b + cells (microglia) from the cell suspension (lower left corner from left panel). The CD11b - cells (bottom half from middle panel) were then sorted into two distinct cell populations corresponding to astrocytes (GLT1 + /CD90.2 - ) and neurons (CD90.2 + /GLT1 - ) (right panel). (D) PCR analysis of astrocytes and neurons from the cell sorting. Microglia (CD11 + ) as well as astrocytes (GLT1 + /CD90.2 - ), but not neurons (CD90.2 + /GLT1 - ) from PHP.eB Gfap-iCre infected animals show the presence of iCre , indicating that only glia and not neurons could express iCre in order to delete Per2 in the Per2 fl/fl mice. (E) Immunohistochemistry of vG Per2 brain tissue from nucleus accumbens (NAc) isolated at ZT6. The signal for PER2 (green) mainly overlaps with neuronal NeuN signal (red) giving rise to the yellow color. Scale bar: 100 µm. (F) Immobility time in the forced swim test (FST) of vG Per2 (PHP.eB Gfap-iCre , green) and control (PHP.eB control, blue) animals are shown (n = 5, two-tailed t-test, *p

    Article Snippet: Antibodies targeted CD11b (fluorophore PECy7, ref. 552850, BD Pharmingen), CD90.2 (fluorophore PE, ref., 130-102-489, Miltenyi Biotech) and GLT1 (fluorophore ATTO 633, ref., AGC-022-FR, Alomone lab), and were used in a 1:100 dilution.

    Techniques: Mouse Assay, Imaging, Injection, Construct, Fluorescence, Flow Cytometry, Polymerase Chain Reaction, FACS, Infection, Immunohistochemistry, Isolation, Two Tailed Test