SDS–PAGE and Western blot analysis were performed exactly as described previously (Barbosa et al., 2015 (link)). Antibodies used were as follows. Primary antibodies were mouse anti-V5, (1:10,000; Life Technologies), rabbit anti–actin 20-36 (1:2500; Sigma-Aldrich), mouse anti–actin AC-40 (1:2500; Sigma-Aldrich), rabbit anti–γ-tubulin (1:2500; Sigma-Aldrich), anti–ApoA-IV 1D6B6 (1:2000; Cell Signaling, Leiden, Netherlands), rabbit phospho-GSK-3α/β(Ser-21/9) D17D2 (1:2000; Cell Signaling), and anti–CREB-H pS73 and pS81 (1:100; Barbosa et al., 2015 (link)). Secondary antibodies were conjugated with horseradish peroxidase (for detection by conventional chemiluminescence) or with Dylight680 or Dylight800 (Fisher Scientific, Lutterworth, United Kingdom) for detection by laser scanning using the LI-COR Odyssey Image system. The latter method was routinely employed for CREB-H detection using the phosphospecific antibodies because it allowed simultaneous analysis, on one blot, of total CREB-H species with the anti-mouse antibody to the epitope tag in one channel and the phosphorylated species with the anti-rabbit antibody to the phosphorylated peptide in a second channel.
Mouse anti actin ac 40
Manufactured by Merck Group
Mouse anti–actin AC-40 is a laboratory reagent used for the detection of actin in biological samples. It is a monoclonal antibody that specifically binds to the actin protein, allowing for its identification and quantification in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti v5, by Thermo Fisher Scientific (1 mentions)
Dylight 800, by Thermo Fisher Scientific (1 mentions)
Dylight 680, by Thermo Fisher Scientific (1 mentions)
Rabbit anti γ tubulin, by Merck Group (1 mentions)
Mouse anti tubulin dm1a, by Merck Group (1 mentions)
Mouse anti gfp gf28r, by Thermo Fisher Scientific (1 mentions)
Mouse anti talin 8d4, by Merck Group (1 mentions)
Gfp trap magnetic agarose, by Proteintech (1 mentions)
Anti gapdh 14c10, by Cell Signaling Technology (1 mentions)
Mouse anti vinculin hvin 1, by Merck Group (1 mentions)
Rp 8 cpt camps, by Biolog (1 mentions)
2 protocols using mouse anti actin ac 40
1
SDS-PAGE and Western Blot Analysis
2
Reagents for Cellular Signaling Analysis
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Rp-8-CPT-cAMPS (8-(4-chlorophenylthio)adenosine-3′,5′-cyclic monophosphorothioate, Rp-isomer) was purchased from either Biolog and Cayman Chemical with no notable differences in efficacy or potency. Reagents for immunofluorescence and immunoblotting were as follows: rabbit anti-paxillin (Y113; Abcam); mouse anti-vinculin (hVin-1; Sigma); rabbit anti-PKA RIIα (sc909; Santa Cruz Biotechnology); mouse anti-talin (8d4; Sigma); PKA-Cα (610980; BD biosciences); Prx3 (PF-PA0255; Abfrontier); rabbit monoclonal anti–phospho-RII (pS99 clone E151, ab32390; Abcam); mouse anti-tubulin (DM1a; Sigma); anti-GAPDH (14C10; Cell Signaling Technologies); mouse anti-actin (AC-40; Sigma); mouse anti-GFP (GF28R; Thermo Fisher); rabbit anti-phospho-PKA substrate (100G7E; Cell Signaling Technologies); and anti-Tns3 antibodies (PA5-63112; Invitrogen and HPA055338; Millipore-Sigma). Secondary reagents were purchased from Jackson ImmunoResearch (Alexa594-and Alexa647-conjugated donkey anti-rabbit and Alexa488-conjugated donkey anti-mouse) and Invitrogen (Alexa488-phalloidin; DAPI (4′,6-diamidino-2-phenylindole)). GFP-Trap Magnetic Agarose was from ProteinTech. Reagents for the synthesis of ATP-biotin were described elsewhere (57 (link)). Unless otherwise noted, other common chemicals and reagents were from Sigma or Fisher.
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