Trichloroacetic Acid Whole Cell Extraction

Whole cell extracts were prepared using a trichloroacetic acid (TCA) method as previously described (11 (link)). Samples were resolved on an 8% or 12% SDS-PAGE gel and transferred onto a PVDF membrane (Immobilon-P; Millipore) using a semi-dry method. Anti-Myc and anti-HA antibody antibodies were purchased from MBL. Anti-FLAG antibody for Western blot was obtained from Sigma (F1804). Anti-mouse and rabbit IgG HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology. Blots were developed using the Western Blotting substrate (Bio-Rad).

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Zheng S., Li D., Lu Z., Liu G., Wang M., Xing P., Wang M., Dong Y., Wang X., Li J., Zhang S., Peng H., Ira G., Li G, & Chen X. (2018). Bre1-dependent H2B ubiquitination promotes homologous recombination by stimulating histone eviction at DNA breaks. Nucleic Acids Research, 46(21), 11326-11339.

Publication 2018

Immobilon-P Anti-mouse and rabbit IgG HRP-conjugated secondary antibodies

Anti antibodies Anti antibody Antibodies Antibody Cell extracts Igg hrp Immobilon p Membrane Mouse Pvdf Rabbit Sds page Trichloroacetic acid

Corresponding Organization : Wuhan University

Other organizations : Institute of Biophysics, Chinese Academy of Sciences, Baylor College of Medicine

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Variable analysis

independent variables

Protein samples prepared using a trichloroacetic acid (TCA) method

dependent variables

Protein expression levels
Protein molecular weights

control variables

SDS-PAGE gel concentration (8% or 12%)
Transfer of proteins onto PVDF membrane using a semi-dry method
Primary antibodies: Anti-Myc, Anti-HA, Anti-FLAG
Secondary antibodies: Anti-mouse and rabbit IgG HRP-conjugated
Detection method: Western Blotting substrate (Bio-Rad)

controls

Positive control: Not specified
Negative control: Not specified

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