ab against map 2  (ProSci Incorporated)


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    ProSci Incorporated ab against map 2
    Rat cerebrocortical cultures were treated with gp120 from HIV-1 strain BaL (200 pM) for 24 h in the presence or absence of different IFNβ concentrations (500 or 5,000 U/ml) or BSA/PBS (0.001% final concentration) as vehicle control. Following the incubation the cells were fixed and permeabilized, and neurons were immunolabeled for neuronal <t>MAP-2</t> and NeuN while nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments with 4–7 replicates and an average of 5,300 cells counted per condition; ***p < 0.001, **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.
    Ab Against Map 2, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury"

    Article Title: IFNβ Protects Neurons from Damage in a Murine Model of HIV-1 Associated Brain Injury

    Journal: Scientific Reports

    doi: 10.1038/srep46514

    Rat cerebrocortical cultures were treated with gp120 from HIV-1 strain BaL (200 pM) for 24 h in the presence or absence of different IFNβ concentrations (500 or 5,000 U/ml) or BSA/PBS (0.001% final concentration) as vehicle control. Following the incubation the cells were fixed and permeabilized, and neurons were immunolabeled for neuronal MAP-2 and NeuN while nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments with 4–7 replicates and an average of 5,300 cells counted per condition; ***p < 0.001, **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.
    Figure Legend Snippet: Rat cerebrocortical cultures were treated with gp120 from HIV-1 strain BaL (200 pM) for 24 h in the presence or absence of different IFNβ concentrations (500 or 5,000 U/ml) or BSA/PBS (0.001% final concentration) as vehicle control. Following the incubation the cells were fixed and permeabilized, and neurons were immunolabeled for neuronal MAP-2 and NeuN while nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments with 4–7 replicates and an average of 5,300 cells counted per condition; ***p < 0.001, **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.

    Techniques Used: Concentration Assay, Incubation, Immunolabeling, Staining, Fluorescence, Microscopy, Cell Counting

    ( a ) Mixed neuronal-glial cerebrocortical cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL (200 pM) and mouse IFNβ (5,000 U/ml) in the presence and absence of neutralizing antibodies against CCL3, CCL4, CCL5, IFNγ or CXCL10. IFNγ antibody was used as control for neutralizing antibodies since this protein was undetectable in cerebrocortical cell cultures. ( b ) Mouse cerebrocortical cultures from IFNAR1KO mice were stimulated with gp120 BaL for 24 h the presence or absence of mouse IFNβ (5,000 U/ml) or BSA/PBS control. ( c ) Cerebrocortical cell cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL in the presence and absence of murine CCL4 (2 or 20 nM). Neuronal survival was assessed by immunofluorescence microscopy and counting of MAP-2/NeuN double-positive neurons. Values are mean ± s.e.m.; n = 3–5 independent experiments with 3–7 replicates and an average of 9,000 (IFNAR1KO) or 5,700 (WT) cells counted per condition; ** p < 0.01, *** p < 0.001 by ANOVA with Fisher’s PLSD post hoc test.
    Figure Legend Snippet: ( a ) Mixed neuronal-glial cerebrocortical cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL (200 pM) and mouse IFNβ (5,000 U/ml) in the presence and absence of neutralizing antibodies against CCL3, CCL4, CCL5, IFNγ or CXCL10. IFNγ antibody was used as control for neutralizing antibodies since this protein was undetectable in cerebrocortical cell cultures. ( b ) Mouse cerebrocortical cultures from IFNAR1KO mice were stimulated with gp120 BaL for 24 h the presence or absence of mouse IFNβ (5,000 U/ml) or BSA/PBS control. ( c ) Cerebrocortical cell cultures from WT mice were simultaneously exposed for 3 days to HIV gp120 BaL in the presence and absence of murine CCL4 (2 or 20 nM). Neuronal survival was assessed by immunofluorescence microscopy and counting of MAP-2/NeuN double-positive neurons. Values are mean ± s.e.m.; n = 3–5 independent experiments with 3–7 replicates and an average of 9,000 (IFNAR1KO) or 5,700 (WT) cells counted per condition; ** p < 0.01, *** p < 0.001 by ANOVA with Fisher’s PLSD post hoc test.

    Techniques Used: Immunofluorescence, Microscopy

    Three to four month-old HIVgp120tg and WT littermate male mice received intranasal recombinant murine IFNβ (50,000 U/25 g bodyweight) or vehicle once a week for four weeks and afterwards, the brains were assessed for neuronal damage and glial activation. Representative images of frontal cerebral cortex ( a ) and hippocampus ( b ) immunolabeled for neuronal synaptophysin (SYP; cortex layer III, hippocampus CA1) deconvolution microscopy; scale bar, 40 μm) and MAP-2 (cortex layer III (area between dashed lines) and hippocampus) fluorescence microscopy; scale bar, 100 μm), astrocytic GFAP (scale bar, 100 μm) and microglial Iba1 (red, DNA in blue, scale bar, 100 μm). ( c–f ) Quantification of microscopy data obtained in frontal cortex and hippocampus of sagittal brain sections of four to five month-old HIVgp120tg mice and controls treated with IFNβ or vehicle: ( c ) SYP in percent positive neuropil, ( d ) MAP-2 immunoreactivity as sum of fluorescence intensity (SFLI; arbitrary units); ( e ) fluorescence signal for astrocytic GFAP; ( f ) quantification of Iba1 + microglia (counts/microscopic field). Values are mean ± s.e.m.; ***p < 0.001, **p < 0.01, *p < 0.05; ANOVA and Fisher’s PLSD post hoc test; n = 4–5 animals per group/genotype.
    Figure Legend Snippet: Three to four month-old HIVgp120tg and WT littermate male mice received intranasal recombinant murine IFNβ (50,000 U/25 g bodyweight) or vehicle once a week for four weeks and afterwards, the brains were assessed for neuronal damage and glial activation. Representative images of frontal cerebral cortex ( a ) and hippocampus ( b ) immunolabeled for neuronal synaptophysin (SYP; cortex layer III, hippocampus CA1) deconvolution microscopy; scale bar, 40 μm) and MAP-2 (cortex layer III (area between dashed lines) and hippocampus) fluorescence microscopy; scale bar, 100 μm), astrocytic GFAP (scale bar, 100 μm) and microglial Iba1 (red, DNA in blue, scale bar, 100 μm). ( c–f ) Quantification of microscopy data obtained in frontal cortex and hippocampus of sagittal brain sections of four to five month-old HIVgp120tg mice and controls treated with IFNβ or vehicle: ( c ) SYP in percent positive neuropil, ( d ) MAP-2 immunoreactivity as sum of fluorescence intensity (SFLI; arbitrary units); ( e ) fluorescence signal for astrocytic GFAP; ( f ) quantification of Iba1 + microglia (counts/microscopic field). Values are mean ± s.e.m.; ***p < 0.001, **p < 0.01, *p < 0.05; ANOVA and Fisher’s PLSD post hoc test; n = 4–5 animals per group/genotype.

    Techniques Used: Recombinant, Activation Assay, Immunolabeling, Microscopy, Fluorescence

    Sagittal brains sections of HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle (veh) were immunolabeled for CCL4, neuronal MAP-2 or astrocytic GFAP. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was labeled with Hoechst (H) 33342. The fluorescence-labeled brain sections were analyzed using confocal laser-scanning microscopy. Representative images of cortex layer III are shown; scale bar, 50 μm.
    Figure Legend Snippet: Sagittal brains sections of HIVgp120tg and WT littermate mice previously treated with intranasal IFNβ or vehicle (veh) were immunolabeled for CCL4, neuronal MAP-2 or astrocytic GFAP. Alexa Fluor 488, 555 and 647 conjugated secondary antibodies were employed to visualize primary Abs and nuclear DNA was labeled with Hoechst (H) 33342. The fluorescence-labeled brain sections were analyzed using confocal laser-scanning microscopy. Representative images of cortex layer III are shown; scale bar, 50 μm.

    Techniques Used: Immunolabeling, Labeling, Fluorescence, Confocal Laser Scanning Microscopy

    ( a ) Cerebrocortical cultures from mice were prepared to either contain microglia, neurons and astrocytes (M + N + A) or were depleted of microglia (N + A) or neurons and microglia (A). Complete and depleted cell cultures were incubated with mIFNβ (5,000 U/ml) or BSA/PBS vehicle control for 0, 3, 6, 12 and 24 h and concentrations of CCL4 were measured in cell-free supernatants using a commercially available multiplex assay as described in Methods. Maximum concentrations were reached in samples of 12 to 24 h mIFNβ exposure and compared to vehicle-treated, baseline samples. Values are mean ± s.e.m.; n = 3 independent experiments; *p < 0.05, student’s t-test. ( b ) Microglia-depleted rat cerebrocortical cultures were exposed for 24 h to 50% cell-free conditioned media (CM) from human MDM in the presence or absence of human IFNβ (5,000 U/ml). MDM were previously stimulated for 24 h with HIV-1 gp120 BaL (MDM gp120 CM) or vehicle (MDM CM). Following the incubation the cells were fixed and permeabilized. Neurons were immunolabeled for neuronal MAP-2 and NeuN and nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments, with 4–8 replicates and an average of 4,000 cells counted per condition; **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.
    Figure Legend Snippet: ( a ) Cerebrocortical cultures from mice were prepared to either contain microglia, neurons and astrocytes (M + N + A) or were depleted of microglia (N + A) or neurons and microglia (A). Complete and depleted cell cultures were incubated with mIFNβ (5,000 U/ml) or BSA/PBS vehicle control for 0, 3, 6, 12 and 24 h and concentrations of CCL4 were measured in cell-free supernatants using a commercially available multiplex assay as described in Methods. Maximum concentrations were reached in samples of 12 to 24 h mIFNβ exposure and compared to vehicle-treated, baseline samples. Values are mean ± s.e.m.; n = 3 independent experiments; *p < 0.05, student’s t-test. ( b ) Microglia-depleted rat cerebrocortical cultures were exposed for 24 h to 50% cell-free conditioned media (CM) from human MDM in the presence or absence of human IFNβ (5,000 U/ml). MDM were previously stimulated for 24 h with HIV-1 gp120 BaL (MDM gp120 CM) or vehicle (MDM CM). Following the incubation the cells were fixed and permeabilized. Neurons were immunolabeled for neuronal MAP-2 and NeuN and nuclear DNA was stained with H33342. Neuronal survival was assessed using fluorescence microscopy and cell counting as described in Methods. Values are mean ± s.e.m.; n = 2 independent experiments, with 4–8 replicates and an average of 4,000 cells counted per condition; **p < 0.01, *p < 0.05 by ANOVA with Fisher’s PLSD post hoc test.

    Techniques Used: Incubation, Multiplex Assay, Immunolabeling, Staining, Fluorescence, Microscopy, Cell Counting

    amino acids 19 34  (ProSci Incorporated)


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    ProSci Incorporated amino acids 19 34
    Amino Acids 19 34, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    amino acids 19 34  (ProSci Incorporated)


    Bioz Verified Symbol ProSci Incorporated is a verified supplier
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    ProSci Incorporated amino acids 19 34
    Amino Acids 19 34, supplied by ProSci Incorporated, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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