Immunofluorescence staining was performed using standard procedures7 (link),71 . Primary antibodies used were mouse anti-Fas III (1:200, DSHB, 7G10), anti-HA (1:200; Sigma-Aldrich H3663), anti-PCNA (1:200; Santa Cruz sc-56), anti-GFP (1:1,000; Abcam ab 13970), anti-mKO (1:200; MBL PM051M), anti-mCherry (1:1000; Invitrogen M11217), anti-H3K27me3 (1:200; Millipore 07–449), anti-H4K20me2/3 (1:400; Abcam ab7817), anti-ssDNA (1:100, DSHB) and anti-BrdU (1:200; Abcam ab6326). BrdU analogue was Invitrogen B23151 5-bromo-2´-deoxyuridine (BrdU). Secondary antibodies were the Alexa Fluor-conjugated series (1:1000; Molecular Probes). Confocal images for immunostained fixed sample were taken using Zeiss LSM 700 Multiphoton confocal microscope with 63x or 100x oil immersion objectives and processed using Adobe Photoshop software.
Sc 56
Manufactured by Abcam
Sc-56 is a monoclonal antibody that recognizes the p56 subunit of the c-Src tyrosine kinase. It can be used for the detection of the c-Src protein in various applications such as Western blotting and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
H3663, by Santa Cruz Biotechnology (2 mentions)
Anti mcherry, by Thermo Fisher Scientific (2 mentions)
M11217, by Merck Group (2 mentions)
Anti h4k20me2 3, by Abcam (2 mentions)
Alexa fluor conjugated series, by Thermo Fisher Scientific (2 mentions)
Lsm 700 multiphoton confocal microscope, by Adobe (2 mentions)
Anti h3k27me3, by Merck Group (2 mentions)
Ab291, by Abcam (1 mentions)
Ab9106, by Santa Cruz Biotechnology (1 mentions)
Ab5467, by Abcam (1 mentions)
Typhoon biomolecular imager, by Cytiva (1 mentions)
3 protocols using sc 56
1
Immunofluorescence Staining of Cellular Markers
2
Immunofluorescence Staining of Cellular Markers
3
Western Blot Analysis of Phospho-H2A and Cell Cycle Markers
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Cells in supplemented EMM were collected and whole-cell protein extract was prepared by vortexing acid-washed glass beads in 20% trichloroacetic acid (TCA) and washing beads with 5% TCA. Lysates were boiled for 5 min in Laemmli Sample buffer (4% SDS, 60 mM Tris-HCl, pH 6.8, 5% glycerol, 5% 2-mercaptoethanol, 0.01% bromophenol blue) and analyzed by SDS-PAGE. Primary antibodies used were as follows: anti-phospho-H2A (Abcam ab17353; 1:1,000), anti-H2A (Cell Signaling 3636S; 1:1,000; re-probed after phospho-H2A), anti-GFP (Abcam ab291; 1:1,000), anti-myc (Abcam ab9106; 1:1,000) anti-PCNA (Santa Cruz sc-56; 1:1,000), and anti-cdc2 (Abcam ab5467; 1:1,000). After secondary antibody (Alexa Flour 488 or 647; 1:6,000) incubation, blots were developed using Amersham Typhoon biomolecular imager. Intensity of bands were quantified with ImageJ.
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