Lysates were generated by incubating cells or ground tumors in RIPA buffer (Millipore). 20–30 μg of cleared lysate were analyzed by SDS-PAGE as previously described 11 (link). The following primary antibodies were used: Vinculin (1:5,000 dilution; Sigma, V9131), β-Actin (1:5,000; Sigma, A5441), Collagen I (1:500; Abcam, ab21286), Fibronectin (1:1,000; Abcam, ab2413), Smad4 (1:200; Santa Cruz, sc-7966), Smad2 p-S465/467 (1:1,000; Cell Signaling, 3108S), Smad2/3 (1:1,000; Cell Signaling, 3102S), GCN2 p-T899 (1:1,000; Abcam, ab75836), GCN2 (1:1,000; Cell Signaling, 3302S), ATF4 (1:200; Santa Cruz, sc-200), PC (1:1,000; Novus, NBP1–49536), S6K (1:1,000; Cell Signaling, 2708S), GLUL (1:1,000; Sigma, G2781), SMA (1:1,000; Millipore, CBL171). The following secondary antibodies were used: anti-rabbit HRP (1:5,000; GE, NA934V), anti-mouse HRP (1:5,000; GE, NA931). Quantification of band intensities of Collagen I relative to β-Actin or Vinculin in tumor allograft experiments was performed with Image Lab software v6.0 (Bio-Rad).
Smad2 p s465 467
Manufactured by Cell Signaling Technology
Smad2 p-S465/467 is a laboratory product that detects the phosphorylation of Smad2 at serine residues 465 and 467. This phosphorylation event is a key step in the activation of the TGF-beta signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Vinculin, by Bio-Rad (2 mentions)
Fibronectin, by Abcam (2 mentions)
Vinculin, by Merck Group (2 mentions)
Smad2 3, by Cell Signaling Technology (2 mentions)
Gcn2 p t899, by Abcam (2 mentions)
Nbp1 49536, by Cell Signaling Technology (2 mentions)
Collagen 1, by Bio-Rad (2 mentions)
Image lab software v 6, by Bio-Rad (2 mentions)
Ripa buffer, by Merck Group (2 mentions)
Smad4, by Santa Cruz Biotechnology (2 mentions)
Collagen 1, by Abcam (2 mentions)
β actin, by Merck Group (2 mentions)
β actin, by Bio-Rad (2 mentions)
2 protocols using smad2 p s465 467
1
Western Blot Analysis of Tumor Proteins
2
Western Blot Analysis of Tumor Proteins
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Lysates were generated by incubating cells or ground tumors in RIPA buffer (Millipore). 20–30 μg of cleared lysate were analyzed by SDS-PAGE as previously described 11 (link). The following primary antibodies were used: Vinculin (1:5,000 dilution; Sigma, V9131), β-Actin (1:5,000; Sigma, A5441), Collagen I (1:500; Abcam, ab21286), Fibronectin (1:1,000; Abcam, ab2413), Smad4 (1:200; Santa Cruz, sc-7966), Smad2 p-S465/467 (1:1,000; Cell Signaling, 3108S), Smad2/3 (1:1,000; Cell Signaling, 3102S), GCN2 p-T899 (1:1,000; Abcam, ab75836), GCN2 (1:1,000; Cell Signaling, 3302S), ATF4 (1:200; Santa Cruz, sc-200), PC (1:1,000; Novus, NBP1–49536), S6K (1:1,000; Cell Signaling, 2708S), GLUL (1:1,000; Sigma, G2781), SMA (1:1,000; Millipore, CBL171). The following secondary antibodies were used: anti-rabbit HRP (1:5,000; GE, NA934V), anti-mouse HRP (1:5,000; GE, NA931). Quantification of band intensities of Collagen I relative to β-Actin or Vinculin in tumor allograft experiments was performed with Image Lab software v6.0 (Bio-Rad).
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