Primary MEFs were seeded in chamber slides, washed with PBS and fixed with 3% PFA/PBS for 12 min at room temperature. Cells were permeabilized with 0.5% Triton solution (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM Sucrose, 0.5% Triton X-100) for 5 min, blocked with 5% normal goat serum for 1 hour and stained with primary antibodies overnight at 4ͦC. Cells were washed with PBS and stained with FITC/TRITC secondary antibodies. Samples were then incubated with DAPI for 1 min, washed with PBS and mounted with ProLong™ Gold Antifade Mountant (Invitrogen). Cells were imaged with Nikon Eclipse Ni-E upright fluorescence microscope. Primary antibodies of HP1α (CST, 2616, 1:200), HP1β (CST, 8676, 1:800), H3 (Immunoway, YM3038, 1:10000), H3K9me3 (Abcam, Ab8898, 1:500), H3K27me3 (CST, 9733, 1:1000), H3K4me3 (Active Motif, 61379,1:500), H4K20me3 (Active Motif, 39671, 1:500), Orc2 (gift from Bruce Stillman, 920–4, 1:200) were used at indicated concentrations for immunofluorescence.
Ym3038
Manufactured by Abcam
YM3038 is a lab equipment product. It is a type of device used in laboratory settings for various research and analytical purposes. The core function of this product is to perform specific tasks related to the handling or processing of samples or materials in a laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ym3038
1
Immunofluorescence Analysis of Chromatin Markers
2
Immunofluorescence Staining of Primary MEFs
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Primary MEFs were seeded in chamber slides, washed with PBS and fixed with 3% PFA/PBS for 12 min at room temperature. Cells were permeabilized with 0.5% Triton solution (20 mM HEPES, 50 mM NaCl, 3 mM MgCl2, 300 mM Sucrose, 0.5% Triton X‐100) for 5 min, blocked with 5% normal goat serum for 1 hr and stained with primary antibodies overnight at 4°C. Cells were washed with PBS and stained with FITC/TRITC secondary antibodies. Samples were then incubated with DAPI for 1 min, washed with PBS and mounted with ProLong™ Gold Antifade Mountant (Invitrogen). Cells were imaged with Nikon Eclipse Ni‐E upright fluorescence microscope. Primary antibodies of HP1α (CST, 2616, 1:200), HP1β (CST, 8676, 1:800), H3 (Immunoway, YM3038, 1:10000), H3K9me3 (Abcam, Ab8898, 1:500), H3K27me3 (CST, 9733, 1:1000), H3K4me3 (Active Motif, 61379,1:500), H4K20me3 (Active Motif, 39,671, 1:500), Orc2 (gift from Bruce Stillman, 920‐4, 1:200) were used at indicated concentrations for immunofluorescence.
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