Protein Expression and Quantification

Cells were lysed in RIPA buffer with protease inhibitors and protein concentration was determined using the BioRad dye-binding protein assay. Equivalent amounts of total protein were analyzed by SDS-PAGE with antibodies specified above, as previously described (McFarland et al. 2013 (link); Rajbhandari et al. 2015 (link)).

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Meares G.P., Rajbhandari R., Gerigk M., Tien C.L., Chang C., Fehling S.C., Rowse A., Mulhern K.C., Nair S., Gray G.K., Berbari N.F., Bredel M., Benveniste E.N, & Nozell S.E. (2018). MicroRNA-31 is Required for Astrocyte Specification. Glia, 66(5), 987-998.

Publication 2018

dye-binding protein assay

Antibodies Assay Binding protein Buffer Cells Protease inhibitors Protein Ripa Sds page

Corresponding Organization : University of Alabama at Birmingham

Other organizations : West Virginia University, Indiana University – Purdue University Indianapolis

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Variable analysis

independent variables

Lysis conditions (RIPA buffer with protease inhibitors)

dependent variables

Protein expression levels (analyzed by SDS-PAGE and antibodies)

control variables

Equivalent amounts of total protein analyzed

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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