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Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and <t>filipin,</t> respectively, n ≥3.
Filipin, supplied by PolyScience, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention

Journal: Human Molecular Genetics

doi: 10.1093/hmg/ddy112

Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and filipin, respectively, n ≥3.
Figure Legend Snippet: Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and filipin, respectively, n ≥3.

Techniques Used: Marker, Expressing, Activation Assay, Fluorescence, Isolation, Mouse Assay, Staining, Labeling

Related Articles

Isolation:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: Paragraph title: Microglia isolation, FACS sorting and analysis ... Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer.

Blocking Assay:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: Microglia at the 70% interphase were collected, washed twice in FACS buffer and stained with fixable LIVE/DEAD fluorescent or pacific blue dye (UV–blue L-23105 or Violet L-34955) for 10 min (1:500) in PBS at 4°C and then incubated with rat anti-mouse CD16/32 (2.4G2; BD Biosciences, San Jose, USA) for 10 min (1:100) in FACS buffer at 4°C to block Fc receptors. .. Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer.

Incubation:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: .. Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer. .. After three washes with FACS buffer, stained cells were fixed using 2% paraformaldehyde prior analysis.

FACS:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: Paragraph title: Microglia isolation, FACS sorting and analysis ... Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer.

Staining:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: Intracellular staining for GM1 and unesterified cholesterol were performed on LIVE/DEAD (violet), CX3 CR1, CD45, CD11b fixed for 20 min using the cytofix/cytoperm buffer [Beckton Dickinson (BD), Franklin Lakes, USA] and washed two times with BD wash buffer. .. Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer.

Software:

Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention
Article Snippet: Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer. .. Stained sample were analyzed using a LSR Fortessa (BD) and the data analyzed using FlowJo software (TreeStar, Ashland, USA).

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    PolyScience filipin
    Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and <t>filipin,</t> respectively, n ≥3.
    Filipin, supplied by PolyScience, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/filipin/product/PolyScience
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    filipin - by Bioz Stars, 2020-04
    93/100 stars
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    Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and filipin, respectively, n ≥3.

    Journal: Human Molecular Genetics

    Article Title: Microglia activation in Niemann–Pick disease, type C1 is amendable to therapeutic intervention

    doi: 10.1093/hmg/ddy112

    Figure Lengend Snippet: Surface marker and biochemical characterization of Npc1 microglial. ( A ) Microglia surface marker expression was characterized in Npc1 +/+ (red) and Npc1 −/− (dashed blue) microglia at birth, 3-, 7- and 9-weeks of age. Microglia steady state markers (CD11b, CD45, CX3CR1 and F4/80) and activation markers (CD86 and MHCII) were quantified. ( B ) GM1 and unesterified cholesterol storage as mean fluorescence intensity (MFI) at birth, 3-, 7- and 9-weeks of age in microglia isolated from Npc1 +/+ (red) and Npc1 −/− (dashed blue) mice. GM1 and unesterified cholesterol accumulation was quantified by staining isolated microglial cells with Alexa488-labeled cholera toxin and filipin, respectively, n ≥3.

    Article Snippet: Filipin 0.1% (m/v) (Polyscience, Warrington, USA) and FITC-conjugated cholera toxin (1/300) were incubated for 2 h at 4°C before being washed twice with BD wash buffer.

    Techniques: Marker, Expressing, Activation Assay, Fluorescence, Isolation, Mouse Assay, Staining, Labeling