Western blot analysis was performed following standard procedures. Whole-cell protein lysates were prepared using RIPA buffer with cOmpleteTM Mini protease inhibitor (Sigma, CN: 11836170001) and PhosSTOP (Sigma, CN: 4906845001). Protein concentrations were assayed using the Pierce BCA assay (ThermoFisher, CN: 23225), then equal amounts of protein were suspended in SDS loading buffer with DTT, separated on Criterion TGX gels 4–15% SDS-PAGE gels (BioRad, CN: 5671084), and transferred onto PVDF membrane using the Transblot Turbo (BioRad) system. The membranes were then treated with appropriate antibodies according to the methods recommended by their manufacturer and LiCor, then imaged on a LiCor CLx imager. The following antibodies were used in this study: ROR1 (1:1000 dilution, CN: D6T8C), BCL2 (1:1000 dilution, CN: 15071) and β-actin (1:1000 dilution, CN: 3700) from Cell Signaling Technologies. Phopho-ROR1 antibody (1:250 dilution) was a gift from Kancera, and was produced as previously described10 (link).
Criterion tgx gels 4 15 sds page gels
Manufactured by Bio-Rad
Criterion TGX gels 4–15% SDS-PAGE gels are pre-cast polyacrylamide gels designed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separation of proteins. These gels have a gradient of 4% to 15% acrylamide concentration, providing a wide separation range for molecular weight analysis of proteins.
Automatically generated - may contain errors
Lab products found in correlation
Trans blot turbo, by Bio-Rad (2 mentions)
Phosstop, by Merck Group (2 mentions)
Pierce bca assay, by Thermo Fisher Scientific (2 mentions)
Subcellular protein fractionation kit, by Thermo Fisher Scientific (2 mentions)
Complete mini protease inhibitor, by Merck Group (1 mentions)
Clx imager, by LI COR (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using criterion tgx gels 4 15 sds page gels
1
Western Blot Analysis of Protein Expression
2
Whole Cell Protein Extraction and Analysis
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