For western blotting, brain samples were lysed by RIPA buffer and loaded onto 8% SDS-PAGE gels and transferred to nitrocellulose membranes (Millipore, MA, USA). Membranes were then incubated in 5% bovine serum albumin for 1 h at room temperature followed by overnight incubation with appropriate primary antibodies (PTEN antibody, sc-7974, Santa Cruz Biotechnology, CA, USA; phosphorylated-AKT (p-AKT) antibody, #9271, Cell Signaling Technology, MA, USA; AKT antibody, sc-8312, Santa Cruz Biotechnology; beta-actin antibody, sc-47778, Santa Cruz Biotechnology). The membranes were then incubated for 1 h at room temperature with secondary antibodies conjugated with HRP (Invitrogen, CA, USA). The HRP signals were visualized by WestSave chemiluminescent detection kit (AbFrontier, South Korea) and went through densitometric analysis on ImageJ software.
Phosphorylated akt p akt antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Phosphorylated-AKT (p-AKT) antibody is a laboratory research tool used to detect and quantify the phosphorylation of the AKT protein. AKT is a serine/threonine protein kinase that plays a crucial role in various cellular processes, including cell growth, proliferation, and survival. The p-AKT antibody specifically recognizes the phosphorylated form of AKT, allowing researchers to study the activation state of this important signaling molecule.
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2 protocols using phosphorylated akt p akt antibody
1
PTEN and AKT Western Blotting
2
Islet Protein Phosphorylation Analysis
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Islet protein (20–35 µg/well) separated by SDS-PAGE was transferred to polyvinylidene fluoride membranes, which were probed with polyclonal rabbit anti–phosphorylated AKT (pAKT) antibody (1:500; Cell Signaling Technology, Danvers, MA), anti-GPRC6A antibody (1:1,000; MyBioSource, San Diego, CA), or anti-MAS1 (1:500; Abcam). Stripped membranes were reprobed with polyclonal rabbit anti-AKT antibody (1:500; Cell Signaling Technology) or anti- β-actin (1:2,000; Sigma-Aldrich). Secondary antibody was goat anti-rabbit IgG horseradish peroxidase (1:50,000; Dako, Carpinteria, CA). Bands were visualized on a FluorChem M system using enhanced chemiluminescence and quantified using ImageJ software.
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