Protocol detail

Western Blot Analysis of Nuclear Proteins

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At experimental endpoints, cells were harvested in 1X Laemmli buffer.
Alternatively, insoluble nuclear fractions were prepared from cells as described
(21 (link)) and boiled in 1X Laemmli buffer.
Lysates in Laemmli buffer were subjected to western blot as described (22 (link)) using primary antibodies (AR SP107,
abcam; AR-441, Santa Cruz, Actin C4, Santa Cruz; Histone H3 ab32356, abcam)
diluted 1:1000 and secondary antibodies diluted 1:10,000.

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Li Y., Yang R., Henzler C., Ho Y., Passow C., Auch B., Carreira S., Rodrigues D.N., Bertan C., Hwang T.H., Quigley D.A., Dang H.X., Morrissey C., Fraser M., Plymate S.R., Maher C.A., Feng F.Y., De Bono J, & Dehm S.M. (2020). Diverse AR Gene Rearrangements Mediate Resistance to Androgen Receptor Inhibitors in Metastatic Prostate Cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(8), 1965-1976.

Publication 2020

AR-441 Actin C4

Actin Antibodies Cells Histone h3 Laemmli buffer Western blot

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Variable analysis

independent variables

Preparation of insoluble nuclear fractions

dependent variables

Protein expression levels measured by Western blot

control variables

Laemmli buffer used for cell lysis
Primary antibodies diluted 1:1000
Secondary antibodies diluted 1:10,000

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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