Immunoblot Analysis of Protein Markers

Protein was extracted as described [52 (link)], quantitated using BCA assay (Pierce) and subjected to immunoblot analysis using anti-Bmi1 (Abcam; Cambridge, MA), anti-CHOP, anti-calreticulin, anti-phospho-EIF2α, anti-β-tubulin (Cell Signaling, Danvers, MA) and anti-Grp78 (Abcam). Secondary antibodies were from Santa Cruz Biotechnology (Dallas, TX). Molecular weight markers (Cat. # 10748010, 5 µL per run, or Cat. #LC5800, 10 µL per run) for immunoblot analyses were from Invitrogen (Grand Island, NY).

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Kraft C.L., Rappaport J.A., Snook A.E., Pattison A.M., Lynch J.P, & Waldman S.A. (2017). GUCY2C maintains intestinal LGR5+ stem cells by opposing ER stress. Oncotarget, 8(61), 102923-102933.

Publication 2017

anti-Bmi1 anti-CHOP anti-calreticulin anti-phospho-EIF2α anti-β-tubulin anti-Grp78

Antibodies Assay Bmi1 Calreticulin Chop Grp78 Immunoblot analyses Molecular markers Protein Tubulin

Corresponding Organization : Thomas Jefferson University

Other organizations : University of Pennsylvania

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Variable analysis

independent variables

Protein extraction method as described in reference [52]

dependent variables

Protein expression levels of Bmi1, CHOP, calreticulin, phospho-EIF2α, and Grp78

control variables

Protein quantitation using BCA assay
Immunoblot analysis using anti-β-tubulin antibody
Molecular weight markers from Invitrogen

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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