Cells were rinsed with ice-cold PBS and lysed in RIPA buffer containing 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1% NP-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM beta-glycerophosphate, 1 mM sodium orthovanadate, and complete protease inhibitor (Roche Applied Science). The cell lysates were incubated for 20 min at 4°C and processed for SDS page and immunoblotting. The antibodies SMO (ab72130; 1:1000; Abcam), IGF1R, EGFR, pAKT (S473), pAKT (T308), AKT (1:1000; Cell Signaling), actin-HRP (1:10,000; Sigma-Aldrich), Flotillin, Caveolin-1 and Transferrin receptor (TF-R, 1:1000; Santa Cruz, TX) were used overnight at 4°C.
Transferrin receptor tf r
Manufactured by Santa Cruz Biotechnology
The Transferrin receptor (TF-R) is a membrane-bound protein that plays a key role in the cellular uptake of iron. It binds to transferrin, a protein that transports iron in the bloodstream, and facilitates the internalization of the iron-transferrin complex into the cell through receptor-mediated endocytosis.
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2 protocols using transferrin receptor tf r
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Protein Extraction and Immunoblotting Protocol
2
Protein Extraction and Immunoblotting Protocol
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Cells were rinsed with ice-cold PBS and lysed in RIPA buffer containing 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1% NP-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM beta-glycerophosphate, 1 mM sodium orthovanadate, and complete protease inhibitor (Roche Applied Science). The cell lysates were incubated for 20 min at 4°C and processed for SDS page and immunoblotting. The antibodies SMO (ab72130; 1:1000; Abcam), IGF1R, EGFR, pAKT (S473), pAKT (T308), AKT (1:1000; Cell Signaling), actin-HRP (1:10,000; Sigma-Aldrich), Flotillin, Caveolin-1 and Transferrin receptor (TF-R, 1:1000; Santa Cruz, TX) were used overnight at 4°C.
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