Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Primer sequences for real-time reverse transcription polymerase chain reaction.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Sequencing

Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Primary mouse retinal ganglion cells isolated with two-step immunopanning. Glial fibrillary acidic protein–labeled cells were detected with green fluorescence ( B ). Syntaxin 1-labeled cells were detected with red fluorescence ( C ). 4',6-diamidino-2-phenylindole nuclear staining was detected with blue fluorescence ( D ). Merge image was constructed ( A ). Scale bars: 100 μm.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Isolation, Labeling, Fluorescence, Staining, Construct

Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Primary mouse retinal ganglion cells isolated by direct magnetic separation. Glial fibrillary acidic protein–labeled cells were detected with green fluorescence ( B ). Syntaxin 1-labeled cells were detected with red fluorescence ( C ). 4',6-diamidino-2-phenylindole nuclear staining was detected with blue fluorescence ( D ). Merge image was constructed ( A ). Scale bars: 100 μm.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Isolation, Labeling, Fluorescence, Staining, Construct

Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Primary mouse retinal ganglion cells isolated by immunopanning-magnetic separation. Glial fibrillary acidic protein–labeled cells were detected with green fluorescence ( B ). Syntaxin 1-labeled cells were detected with red fluorescence ( C ). 4',6-diamidino-2-phenylindole nuclear staining was detected with blue fluorescence ( D ). Merge image was constructed ( A ). Scale bars: 100 μm.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Isolation, Labeling, Fluorescence, Staining, Construct

Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Representative western immunoblots for glial fibrillary acidic protein, syntaxin 1, and β-actin. Primary mouse retinal ganglion cells were isolated using three different systems, including two-step immunopanning, direct magnetic separation, and immunopanning-magnetic separation.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Western Blot, Isolation

Journal: Molecular Vision

Article Title: Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation

doi:

Figure Lengend Snippet: Quantitative data of real-time reverse transcription polymerase chain reaction (RT-PCR) for glial fibrillary acidic protein (GFAP) and syntaxin 1. A relative RNA ratio was calculated by dividing the value of each purification method by the value of whole retinal cell suspension. Data were expressed as the mean±SEM. (n=9 for each method). The p value for GFAP and syntaxin 1 was 0.021* and 0.022**, respectively.

Article Snippet: Twenty-four hours after seeding, cells were fixed with 4% paraformaldehyde for 30 min, treated with 0.1% Triton X-100 (Sigma-Aldrich, St. Louis, MO) in 0.1% Na-Citrate (Sigma-Aldrich) for 10 min, and then blocked with 2% bovine serum albumin (Sigma-Aldrich) for 1 h. Cells were incubated with astrocyte-specific antimouse glial fibrillary acidic protein (GFAP) antibody (1:100 dilution; Abcam) and amacrine-specific antimouse syntaxin 1 antibody (1:50 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Purification