Western Blot Analysis of CCA Cell Proteins

Western blot analysis of total protein in cell lysates from cultured rat and/or human CCA cell lines was carried out as previously described (17 (link), 20 (link)), with the following primary antibodies purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA): S1PR2 (H64), S1PR3 (H70), ERK1 (C16), ERK-2 (C-14), p-ERK (E-4), Akt1/2/3/ (H-136) and p-Akt1/2/3/ (ser473). S1PR1 (Ab23695) was purchased from Abcam (Cambridge, MA). Protein bands were normalized against β-actin, detected with mouse monoclonal anti-actin antibody (ACTN05) from Thermo Scientific (Wilmington, DE).

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Liu R., Zhao R., Zhou X., Liang X., Campbell D.J., Zhang X., Zhang L., Shi R., Wang G., Pandak W.M., Sirica A.E., Hylemon P.B, & Zhou H. (2014). Conjugated bile acids promote cholangiocarcinoma cell invasive growth through activation of sphingosine 1-phosphate receptor 2. Hepatology (Baltimore, Md.), 60(3), 908-918.

Publication 2014

S1PR2 (H64), S1PR3 (H70), ERK1 (C16), ERK-2 (C-14), p-ERK (E-4), Akt1/2/3/ (H-136) p-Akt1/2/3/ (ser473). S1PR1 (Ab23695)

Actin Akt1 Anti antibody Antibodies Cell Cell lines Erk 2 Erk1 Human Mouse Protein Western blot analysis

Corresponding Organization : Wenzhou Medical University

Other organizations : China Pharmaceutical University, Nanjing Medical University, Jiangsu Province Hospital

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Variable analysis

independent variables

Rat and/or human CCA cell lines

dependent variables

Total protein levels in cell lysates
Protein bands normalized against β-actin

control variables

Western blot analysis protocol as previously described (17, 20)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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