Protein assays (Pierce, Rockford, IL) and Western blot analysis (Bio-Rad, Hercules, CA) were performed as previously described (Cantara et al., 2012 (link)) with 8 μg of protein loaded/well and protein bands quantified using ImageJ (version 1.46r) with normalization to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Levels of each phosphorylated protein were additionally normalized to total protein levels. Antibodies, their phosphorylated epitope (when relevant), their dilution from stock solution, and their source are as follows: GAPDH (1:10,000; Fitzgerald, Acton, MA), p25/TPPP (clone EPR 3316, 1:1000; Epitomics), phospho–LIMK 1 (Thr-508; 1:500; Abcam), phospho–LIMK 2 (T505; 1:500; Abcam), phospho-cofilin (Ser-3; 77G2, 1:1000; Cell Signaling), cofilin (1:2000; Sigma-Aldrich), phospho-FAK (Tyr-576/577; 1:1000; Cell Signaling), phospho-FAK (1:1000; Tyr-397; Cell Signaling), FAK (1:1000; Cell Signaling), phospho-paxillin (Tyr-118; 1:1000; Cell Signaling), paxillin (1:500; Cell Signaling), phospho–histone H3 (pHH3; Ser-10; 1:1000; Cell Signaling), and histone H3 (1:1000; Cell Signaling).
Phospho cofilin ser 3 77g2
Manufactured by Cell Signaling Technology
Phospho-cofilin (Ser-3; 77G2) is a primary antibody that detects endogenous levels of cofilin only when phosphorylated at serine 3. Cofilin is an actin-binding protein that plays a key role in the regulation of actin filament dynamics.
Automatically generated - may contain errors
Lab products found in correlation
Paxillin, by Cell Signaling Technology (1 mentions)
Cofilin, by Merck Group (1 mentions)
Phospho fak, by Cell Signaling Technology (1 mentions)
Gapdh, by Bio-Rad (1 mentions)
Western blot analysis, by Bio-Rad (1 mentions)
Phospho limk 1, by Abcam (1 mentions)
Phospho fak tyr576 577, by Cell Signaling Technology (1 mentions)
Phospho paxillin, by Cell Signaling Technology (1 mentions)
Phospho histone h3 phh3 ser 10, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Gapdh, by Biosynth (1 mentions)
Nb100 122, by Novus Biologicals (1 mentions)
Nb100 2219, by Novus Biologicals (1 mentions)
Anti β tubulin, by Merck Group (1 mentions)
Nb100 479, by Novus Biologicals (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
2 protocols using phospho cofilin ser 3 77g2
1
Quantification of Signaling Proteins
2
Evaluating Hypoxia Signaling Pathways
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Lung tissue samples from mice were homogenized in RIPA lysis buffer supplemented with protease inhibitor cocktail (P8340, Sigma-Aldrich). Western blot analysis was performed using anti-PHD2 (NB100-2219 Novus Biologicals and #4835S Cell Signaling), HIF1α (NB100-479 Novus Biologicals) and HIF2α (NB100-122 Novus Biologicals) antibodies. Anti-β-actin (A2066 Sigma-Aldrich) and anti-β-tubulin antibodies (Sigma T4026) were used as a loading control. Phospho-MYPT1 (Thr696, 5163T), MYPT1 (2634T), phospho-myosin light chain 2 (Thr18/Ser19, 3674T), myosin light chain 2 (D18E2, 8505), phospho-Cofilin (Ser3, 77G2) and Cofilin (D3F9, XP® 5175T) antibodies were all from Cell Signaling Technology.
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