Immunoblotting was conducted using protocols from our previous studies (Gao et al. 2015 (link); Liu and Gao 2020 (link); Liu et al. 2021 (link)). Multi-lane blots were used to confirm the protein changes in animal tissues as observed in 2D-DIGE and probe for the protein changes in NNK transformed BEAS-2B cells. In addition, traditional Western blot was used to measure the protein changes of interest. Calregulin (sc-166837), ezrin (sc-398542), E-cadherin (sc-52328), HRF (sc-133131), and PPA1 (sc-377081) primary antibodies and anti-mouse secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). Vimentin (5741) primary and anti-rabbit (7074) secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA).
Protocol detail
Protein Expression Analysis in Lung Cancer
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2d dige
Animal
Anti antibodies
Antibodies
Calregulin
Cells
E cadherin
Ezrin
Mouse
Protein
Rabbit
Tissues
Vimentin
Western blot
Corresponding Organization :
Other organizations : West Virginia University
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Variable analysis
independent variables
- NNK (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone) transformed BEAS-2B cells
- Protein changes in animal tissues as observed in 2D-DIGE
- Protein changes in NNK transformed BEAS-2B cells
- Protein levels of calregulin, ezrin, E-cadherin, HRF, and PPA1
- Experimental protocols from previous studies (Gao et al. 2015, Liu and Gao 2020, Liu et al. 2021)
- Antibodies used: calregulin (sc-166837), ezrin (sc-398542), E-cadherin (sc-52328), HRF (sc-133131), PPA1 (sc-377081), vimentin (5741)
- Secondary antibodies: anti-mouse, anti-rabbit
- Not explicitly mentioned
- Not explicitly mentioned
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