Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Anti-Histone H3 (Acetyl Lys14) Antibody - http://www.stjohnslabs.com/histone-h3-acetyl-lys14-antibody?filter_name=Stj97144 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation

1. Gel electrophoresis information：10% SDS-PAGE, constant voltage 170V for 60 min
Transfer information：0.45uM nitrocellulose membrane, wet transfer at 250 mA for 60 min.
Lane No. Antigen
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Target band KD
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time
1 MCF7 lysate 60 ug
Histone H3
(Acetyle
Lys14) anti-
body
1:500 1:2500 ~15KD
ECL 7
min
Cell Line: MCF7 cell line
Method of validation: Western Blot
Primary Antibody:
Histone H3 (Acetyle Lys14)
antibody
Secondary Antibody Goat anti rabbit IgG HRP
Dilution ratio: 1:500
Protocol
Treatment of materials: Cells were cultured to 80-90% confluency, and lysed in RIPA buffer.
E.g. Gel electrophoresis: 10% SDS-PAGE, constant voltage 170V for 60 min
Transfer: 0.45uM nitrocellulose membrane, wet transfer at 250 mA for 60 min.
Blocking: 5% milk in TBS for 15 min at RT.
Primary antibody probing: 1/500 diluted in 5% milk in TBS, incubated at 4 C overnight.
Membrane wash: TBS wash 3 times, 5 min/each
Secondary antibody probing: 1/2500 diluted in 5% milk in TBS, incubated at RT for 1 h.
Membrane wash: TBS wash 3 times, 5 min/each
Visualization: ECL 7 min
Antibody Customer Review:
STJ97144 Histone H3 (Acetyle Lys14) antibody
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