Whole cell lysates were prepared in RIPA buffer containing protease inhibitors (Santa Cruz) and quantified using the BCA Protein Assay (Thermo Scientific). Equal amounts of protein were loaded and separated on pre-cast Novex 4–20% SDS–PAGE midi gels (Life Technologies, Carlsbad, CA, USA), transferred to nitrocellulose membranes, and probed with antibodies against SK1 (12071), p53 (9282), p21waf1/cip1 (2947), and pERM (3726) from Cell Signaling Technology (Danvers, MA, USA); SK2 (ab37977) from Abcam; and Actin (A5441) from Sigma. Secondary HRP-conjugate Abs were purchased from Jackson Laboratories (Bar Harbor, ME, USA). Protein bands were revealed using Enhanced Chemiluminescence reagent (Thermo Scientific).
Novex 4 20 sds page midi gels
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Novex 4–20% SDS–PAGE midi gels are precast polyacrylamide gels designed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. They are available in a midi format and have a gradient of 4% to 20% acrylamide concentration.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Santa Cruz Biotechnology (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Actin a5441, by Merck Group (1 mentions)
Enhanced chemiluminescence reagent, by Thermo Fisher Scientific (1 mentions)
Enhanced chemiluminescence ecl reagent, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
β actin, by Merck Group (1 mentions)
β actin, by Thermo Fisher Scientific (1 mentions)
2 protocols using novex 4 20 sds page midi gels
1
Western Blot Analysis of Signaling Proteins
2
Quantitative Western Blot Analysis
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Whole cell lysates were prepared in RIPA buffer containing protease inhibitors (Santa Cruz), and protein concentrations were quantified using the BCA Protein Assay (Thermo Scientific). Equal amounts of proteins were loaded and separated on precast Novex 4–20% SDS–PAGE midi gels (Life Technologies, Carlsbad, CA), transferred to nitrocellulose membranes, and probed with antibodies against SK1 (12071) and PHGDH (13428S) from Cell Signaling Technology (Danvers, MA); and β-actin (A5441) from Sigma. HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). Protein bands were revealed using Enhanced Chemiluminescence (ECL) reagent (Thermo Scientific), and their densities were measured by ImageJ. The density ratio of a protein of interest (POI) to the sample loading control, β-actin, is shown underneath the band of the POI.
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