Stable A3G-expressing cells, stable A3G knockdown cells and control cells were collected, followed by the addition of agarose-conjugated antibodies. Twenty microliters of anti-Akt antibody beads (Cell Signaling Technology, Danvers, MA, USA) was added to 200 μl of the cell lysate and incubated overnight at 4 °C with gentle rocking. The immunoprecipitates were washed twice with 500 μl of 1× cell lysis buffer and twice with 500 μl of 1× kinase buffer. The immunoprecipitates were resuspended in 50 μl of 1× kinase buffer mixed with 200 μM ATP and 1.5 μl of GSK-3 fusion protein and then incubated at 30 °C for 30 min. The mixtures were subjected to western blot analysis with phospho-GSK3β antibody (1:1000, Cell Signaling Technology, Danvers, MA, USA).
Phospho gsk3β antibody
Manufactured by Cell Signaling Technology
Sourced in United States, Japan
The Phospho-GSK3β antibody is a laboratory reagent used in research applications. It is designed to detect and bind to the phosphorylated form of the glycogen synthase kinase 3 beta (GSK3β) protein, which plays a key role in various cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Valproic acid, by Merck Group (1 mentions)
Nestin, by Merck Group (1 mentions)
Trichostatin a, by Merck Group (1 mentions)
Taq polymerase, by Takara Bio (1 mentions)
Protein g agarose, by Merck Group (1 mentions)
Neurod1, by Abcam (1 mentions)
Vglut1, by Abcam (1 mentions)
Mash1, by Abcam (1 mentions)
Synaptophysin antibody, by Abcam (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Ecl reagent, by Cytiva (1 mentions)
Hdac1, by Cell Signaling Technology (1 mentions)
Acetyl histone h3, by Cell Signaling Technology (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
Psd95, by Abcam (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
β actin antibody, by Merck Group (1 mentions)
Glial fibrillary acidic protein (gfap), by Merck Group (1 mentions)
Tuj 1 antibody, by Fortrea (1 mentions)
3 protocols using phospho gsk3β antibody
1
Akt Kinase Activity Assay
2
Neuronal Differentiation Protocols
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The materials used in this study's experiments are listed below along with their respective suppliers: Lithium chloride, Sodium butyrate, Trichostatin A, and Valproic acid from Sigma (St. Louis, MO); ECL™ reagents from Amersham Life Science (Arlington Heights, IL); Trizol® reagent, SuperScript™II Reverse Transcriptase, Tert siRNA [TERT-RSS313560 (rat), TERT-MSS211505-7 (mouse)] from Invitrogen (Carlsbad, CA); Protein G Agarose from Millipore (Billerica, MA); TDZD-8 (4-Benzyl-2-methyl-1,2,4,-Thiadiazolidine-3,5-dione) from Calbiochem (La Jolla, CA); Taq polymerase from Takara (Shiga, Japan).
Antibodies from the following companies: β-actin antibody from Sigma (St. Louis, MO); Tuj-1 antibody from Covance (Princeton, NJ); TERT antibody from Santa Cruz Biotechnology (Santa Cruz, CA); vGluT1, PSD-95, α-CaMKII, NeuroD1, Mash1, BRG1, Synaptophysin antibody from Abcam (Cambridgeshire, England); GAD, Nestin, GFAP, Pax6 and Ngn2GFAP antibody from Millipore (Billerica, MA); HDAC1, Histone H3, Acetyl-Histone H3, GSK3β and phospho-GSK3β antibody from Cell signaling (Boston, MA).
Antibodies from the following companies: β-actin antibody from Sigma (St. Louis, MO); Tuj-1 antibody from Covance (Princeton, NJ); TERT antibody from Santa Cruz Biotechnology (Santa Cruz, CA); vGluT1, PSD-95, α-CaMKII, NeuroD1, Mash1, BRG1, Synaptophysin antibody from Abcam (Cambridgeshire, England); GAD, Nestin, GFAP, Pax6 and Ngn2GFAP antibody from Millipore (Billerica, MA); HDAC1, Histone H3, Acetyl-Histone H3, GSK3β and phospho-GSK3β antibody from Cell signaling (Boston, MA).
3
Salidroside and Donepezil Modulate Tau Phosphorylation
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Salidroside (Sal, Purity > 99.7%) was obtained from the Green Valley Pharmaceutical Corporation (Shanghai, China). It was dissolved in PBS to a stock concentration of 100 mM and stored at − 20 °C. Donepezil was supplied by Eisai Pharmaceutical Co., Ltd. (Tokyo, Japan).
The following antibodies were used: Phospho-GSK-3β antibody, GSK-3β antibody, Mouse monoclonal Phospho-tau (ser396) antibody and tau (Cell Signaling Technology), Mouse monoclonal anti-β-actin antibody (Sigma–Aldrich, Clone AC-15), HRP-conjugated goat anti-mouse IgG (Jackson Immuno Research Laboratories, PA, USA). All chemicals were purchased from Sigma-Aldrich except those noted otherwise.
The following antibodies were used: Phospho-GSK-3β antibody, GSK-3β antibody, Mouse monoclonal Phospho-tau (ser396) antibody and tau (Cell Signaling Technology), Mouse monoclonal anti-β-actin antibody (Sigma–Aldrich, Clone AC-15), HRP-conjugated goat anti-mouse IgG (Jackson Immuno Research Laboratories, PA, USA). All chemicals were purchased from Sigma-Aldrich except those noted otherwise.
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