SDS-PAGE and Protein Immunoblotting

SDS-PAGE was performed as described [22 (link)]. The following antibodies were used: rabbit anti-Na_v1.5 (1:1000; Cell Signaling Technology); rabbit anti-E-cadherin (1:1000; Cell Signaling Technology); rabbit anti-N-cadherin (1:1000; Cell Signaling Technology); rabbit anti-vimentin (1:1000; Cell Signaling Technology); rabbit anti-Slug (1:1000; Cell Signaling Technology); rabbit anti-Snail (1:1000; Cell Signaling Technology); mouse anti-CD44 (1:1000; AbD Serotec); rabbit anti-heat shock protein 90 (HSP90; 1:1000; Cell Signaling Technology), and mouse anti-α-tubulin (1:10,000; Sigma).

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Nelson M., Yang M., Millican-Slater R, & Brackenbury W.J. (2015). Nav1.5 regulates breast tumor growth and metastatic dissemination in vivo. Oncotarget, 6(32), 32914-32929.

Publication 2015

rabbit anti-E-cadherin rabbit anti-N-cadherin rabbit anti-vimentin rabbit anti-Slug rabbit anti-Snail mouse anti-CD44 mouse anti-α-tubulin

Antibodies Cadherin Cadherin 1 Cd44 Hsp90 Mouse Rabbit Sds page Slug Snail Vimentin α tubulin

Corresponding Organization :

Other organizations : University of York, St James's University Hospital

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Variable analysis

independent variables

SDS-PAGE protocol

dependent variables

Protein expression levels of Na_v1.5, E-cadherin, N-cadherin, vimentin, Slug, Snail, CD44, and heat shock protein 90 (HSP90)

control variables

Antibody concentrations: rabbit anti-Na_v1.5 (1:1000), rabbit anti-E-cadherin (1:1000), rabbit anti-N-cadherin (1:1000), rabbit anti-vimentin (1:1000), rabbit anti-Slug (1:1000), rabbit anti-Snail (1:1000), mouse anti-CD44 (1:1000), rabbit anti-heat shock protein 90 (HSP90; 1:1000), and mouse anti-α-tubulin (1:10,000)

controls

No positive controls were explicitly mentioned.
No negative controls were explicitly mentioned.

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