C2C12 myoblasts (ATCC) and the p300 shRNA knockdown cells [36 ] were maintained in proliferating conditions at a confluency lower than 70% in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS (GM) at 37ºC with 5% CO2. Myoblasts were induced to differentiate at about 80% confluence using differentiation medium (DM, DMEM medium containing 2% horse serum). Whole cell extracts were prepared for protein immunoblotting, as described previously [53 ]. Scion Image Software (Scion Corp.) was used for protein band quantification, where band intensities were measured and corrected for against the background of the images. The antibodies used in this study for H4K8ac, H3K9ac, H3K18ac, and H3K27ac were obtained from Abcam (ab15823, ab4441, ab1191, ab4729, respectively), while anti-p300 and anti-MyoD were from Santa Cruz (sc-584x, sc-32758x, respectively). Myogenin antibody was from hybridoma F5D and anti-tubulin from hybridoma E7 [54 ].
Ab4441
Manufactured by Santa Cruz Biotechnology
Ab4441 is a laboratory equipment product offered by Santa Cruz Biotechnology. It is a high-quality research tool designed for specific applications. The core function of Ab4441 is to facilitate critical research processes, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab4729, by Santa Cruz Biotechnology (1 mentions)
Ab1191, by Santa Cruz Biotechnology (1 mentions)
Ab15823, by Abcam (1 mentions)
Sc 584x, by Santa Cruz Biotechnology (1 mentions)
Sc 32758x, by Santa Cruz Biotechnology (1 mentions)
Scion image software, by Techcomp Instruments (1 mentions)
Ultrastick ultrafrost adhesion slides, by Electron Microscopy Sciences (1 mentions)
G9023, by Merck Group (1 mentions)
T9026, by Abcam (1 mentions)
Fluoromount, by Merck Group (1 mentions)
2 protocols using ab4441
1
Investigating Myoblast Differentiation and Epigenetic Regulation
2
Immunofluorescence of Meiotic Germ Cells
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Sorted germ cells and SPZEJ were processed as described previously7 (link),38 (link) and attached to UltraStick/UltraFrost Adhesion slides (Electron Microscopy Sciences). Samples were fixed in 4% PFA in PBS for 15 min before antigen retrieval in three consecutive 5-min incubations with boiling citrate (10 mM at pH 6), followed by triton X-100 (0.2% in PBS) for 15 min. After blocking for one hour in PBST with 5% normal goat serum (Merck G9023) and 0.1% BSA, antibodies were applied overnight at 4 °C in a humidified chamber. The primary antibodies were α-tubulin (Merck T9026; 1:1000), H3K9ac (Abcam ab4441; 1:1000), and Spo11 (Santa Cruz Biotechnology sc-33146; 1:1000). Anti-mouse or anti-rabbit IgG coupled with Alexa-555 (Invitrogen A-21422 and Merck AP510C, respectively) were applied for 1 h at room temperature and cells were counterstained with 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI; Merck G8294; 1:3000) before mounting with Fluoromount™.
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