Staining:Article Title: Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
Article Snippet: .. Glass Coverslips 24 x 50 mm, No.1.5 (Globe scientific inc ., catalog number: 1415-15) Forceps Humidified chamber (Biolegend, catalog number: SIG-31031) Coplin Jar Slide Staining Dish (IHC WORLD, catalog number: IW-2511) Slide Staining Rack (IHC WORLD, catalog number: IW-2512) Xylene (Fisher Scientific, catalog numbers: HC7001GAL, 22-110-676 and X16-4) Isopropanol (Fisher Scientific, catalog number: A416-4) EnVision FLEX Target Retrieval Solution, High pH (50x) (Dako Omnis, Code No: GV804) Tris-HCl (Sigma-Aldrich, catalog number: T5941) NaCl (Sigma-Aldrich, catalog number: S9888) Tween 20 (Sigma-Aldrich, catalog number: BP337) Triton X-100 (Sigma-Aldrich, catalog number: X100) NaN 3 (Fisher Scientific, catalog number: S2271-25) Normal Goat Serum (Sigma-Aldrich, catalog number: G9023) Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number: A7030) Tris Base (Fisher Scientific, catalog number: BP152-5) Anti- γ-tubulin (1:100, Sigma-Aldrich, catalog number: T5326) Anti-Pericentrin (1:200, Abcam, catalog number: 28144) Anti-CEP135 (1:50, Abcam, catalog number: ab75005) Anti-Centrin-3 (1:100, Abnova, catalog number: H00001070-M01) Anti-CP110 (1:100, proteintech TM , catalog number: 12780-I-AP) Anti-E-Cadherin (1:100, Abcam, catalog number: ab76055 M168) Secondary antibodies, purchased from Jackson Immunoresearch and used at 1:200 Hoechst 33342 (1:1,000; Invitrogen, catalog number: H3570) ProLong ® Diamond Antifade Mountant (Thermo Fisher Scientific, catalog number: {"type":"entrez-protein","attrs":{"text":"P36970","term_id":"172045845","term_text":"P36970"}} P36970 ) EDTA (Fisher Scientific, catalog number: BP120-500) Washing buffer for FFPE tissue (see Recipes) Blocking buffer for FFPE tissue (see Recipes) Antigen retrieval buffer (see Recipes) Tris-EDTA Buffer for Antigen Retrieval (pH 9.0) (see Recipes) .. Hot air oven (Labnet ProBlot TM 6 Hybridization Oven) Fume hood Decloaking chamber or pressure cooker (Biocare medical, DC0228) High-resolution fluorescence microscope (DeltaVision Core system (GE Healthcare Bio-Sciences, Pittsburgh, United States of America) equipped with Olympus IX71 Camera (CoolSNAP HQ2; Photometrics) Note: Because centrioles are small (0.5 μm in length) and can localize at close proximity depending on cell cycle, high resolution images at a resolution around 200 nm are required to accurately separate each centrosome.
Formalin-fixed Paraffin-Embedded:Article Title: Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
Article Snippet: .. Glass Coverslips 24 x 50 mm, No.1.5 (Globe scientific inc ., catalog number: 1415-15) Forceps Humidified chamber (Biolegend, catalog number: SIG-31031) Coplin Jar Slide Staining Dish (IHC WORLD, catalog number: IW-2511) Slide Staining Rack (IHC WORLD, catalog number: IW-2512) Xylene (Fisher Scientific, catalog numbers: HC7001GAL, 22-110-676 and X16-4) Isopropanol (Fisher Scientific, catalog number: A416-4) EnVision FLEX Target Retrieval Solution, High pH (50x) (Dako Omnis, Code No: GV804) Tris-HCl (Sigma-Aldrich, catalog number: T5941) NaCl (Sigma-Aldrich, catalog number: S9888) Tween 20 (Sigma-Aldrich, catalog number: BP337) Triton X-100 (Sigma-Aldrich, catalog number: X100) NaN 3 (Fisher Scientific, catalog number: S2271-25) Normal Goat Serum (Sigma-Aldrich, catalog number: G9023) Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number: A7030) Tris Base (Fisher Scientific, catalog number: BP152-5) Anti- γ-tubulin (1:100, Sigma-Aldrich, catalog number: T5326) Anti-Pericentrin (1:200, Abcam, catalog number: 28144) Anti-CEP135 (1:50, Abcam, catalog number: ab75005) Anti-Centrin-3 (1:100, Abnova, catalog number: H00001070-M01) Anti-CP110 (1:100, proteintech TM , catalog number: 12780-I-AP) Anti-E-Cadherin (1:100, Abcam, catalog number: ab76055 M168) Secondary antibodies, purchased from Jackson Immunoresearch and used at 1:200 Hoechst 33342 (1:1,000; Invitrogen, catalog number: H3570) ProLong ® Diamond Antifade Mountant (Thermo Fisher Scientific, catalog number: {"type":"entrez-protein","attrs":{"text":"P36970","term_id":"172045845","term_text":"P36970"}} P36970 ) EDTA (Fisher Scientific, catalog number: BP120-500) Washing buffer for FFPE tissue (see Recipes) Blocking buffer for FFPE tissue (see Recipes) Antigen retrieval buffer (see Recipes) Tris-EDTA Buffer for Antigen Retrieval (pH 9.0) (see Recipes) .. Hot air oven (Labnet ProBlot TM 6 Hybridization Oven) Fume hood Decloaking chamber or pressure cooker (Biocare medical, DC0228) High-resolution fluorescence microscope (DeltaVision Core system (GE Healthcare Bio-Sciences, Pittsburgh, United States of America) equipped with Olympus IX71 Camera (CoolSNAP HQ2; Photometrics) Note: Because centrioles are small (0.5 μm in length) and can localize at close proximity depending on cell cycle, high resolution images at a resolution around 200 nm are required to accurately separate each centrosome.
Blocking Assay:Article Title: Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
Article Snippet: .. Glass Coverslips 24 x 50 mm, No.1.5 (Globe scientific inc ., catalog number: 1415-15) Forceps Humidified chamber (Biolegend, catalog number: SIG-31031) Coplin Jar Slide Staining Dish (IHC WORLD, catalog number: IW-2511) Slide Staining Rack (IHC WORLD, catalog number: IW-2512) Xylene (Fisher Scientific, catalog numbers: HC7001GAL, 22-110-676 and X16-4) Isopropanol (Fisher Scientific, catalog number: A416-4) EnVision FLEX Target Retrieval Solution, High pH (50x) (Dako Omnis, Code No: GV804) Tris-HCl (Sigma-Aldrich, catalog number: T5941) NaCl (Sigma-Aldrich, catalog number: S9888) Tween 20 (Sigma-Aldrich, catalog number: BP337) Triton X-100 (Sigma-Aldrich, catalog number: X100) NaN 3 (Fisher Scientific, catalog number: S2271-25) Normal Goat Serum (Sigma-Aldrich, catalog number: G9023) Bovine serum albumin (BSA) (Sigma-Aldrich, catalog number: A7030) Tris Base (Fisher Scientific, catalog number: BP152-5) Anti- γ-tubulin (1:100, Sigma-Aldrich, catalog number: T5326) Anti-Pericentrin (1:200, Abcam, catalog number: 28144) Anti-CEP135 (1:50, Abcam, catalog number: ab75005) Anti-Centrin-3 (1:100, Abnova, catalog number: H00001070-M01) Anti-CP110 (1:100, proteintech TM , catalog number: 12780-I-AP) Anti-E-Cadherin (1:100, Abcam, catalog number: ab76055 M168) Secondary antibodies, purchased from Jackson Immunoresearch and used at 1:200 Hoechst 33342 (1:1,000; Invitrogen, catalog number: H3570) ProLong ® Diamond Antifade Mountant (Thermo Fisher Scientific, catalog number: {"type":"entrez-protein","attrs":{"text":"P36970","term_id":"172045845","term_text":"P36970"}} P36970 ) EDTA (Fisher Scientific, catalog number: BP120-500) Washing buffer for FFPE tissue (see Recipes) Blocking buffer for FFPE tissue (see Recipes) Antigen retrieval buffer (see Recipes) Tris-EDTA Buffer for Antigen Retrieval (pH 9.0) (see Recipes) .. Hot air oven (Labnet ProBlot TM 6 Hybridization Oven) Fume hood Decloaking chamber or pressure cooker (Biocare medical, DC0228) High-resolution fluorescence microscope (DeltaVision Core system (GE Healthcare Bio-Sciences, Pittsburgh, United States of America) equipped with Olympus IX71 Camera (CoolSNAP HQ2; Photometrics) Note: Because centrioles are small (0.5 μm in length) and can localize at close proximity depending on cell cycle, high resolution images at a resolution around 200 nm are required to accurately separate each centrosome.
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