Pgsk 3α ser21

Manufactured by Cell Signaling Technology

PGSK-3α ser21 is a recombinant protein that corresponds to the phosphorylated form of glycogen synthase kinase 3 alpha (GSK-3α) at serine 21. This protein is useful for research purposes, particularly in studies involving the regulation and signaling pathways of GSK-3α.

Automatically generated - may contain errors

Lab products found in correlation

P gsk3β ser9, by Cell Signaling Technology (2 mentions) P akt ser473, by Cell Signaling Technology (1 mentions) α tubulin, by Merck Group (1 mentions) Pan akt, by Cell Signaling Technology (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Gsk3α, by Cell Signaling Technology (1 mentions) Gsk3β, by Cell Signaling Technology (1 mentions) Lsm 780, by Zeiss (1 mentions)

3 protocols using pgsk 3α ser21

Antibody Analysis of GBM Cell Lines

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The following antibodies were from Cell Signaling (Danvers, MA): pan-AKT(#4685), AKT1(#2938), pAKT Ser-473(#9271, #4060), PARP (#9542), GSK3α (#9338), pGSK-3α ser21 (#9316), GSK3β (#9315), pGSK3β ser9 (#9323), pS6 ribosomal protein ser240/244 (#5364) all used at 1:1000 dilution. Antibodies against α-tubulin (#05-829) and AKT3 (#07-383) were from Millipore (Billerica, MA) and were used at 1 μg/ml and 1:500, respectively.
The GBM cell lines, U87 and U251, were obtained from ATTC (Manassas, VA) and maintained in DMEM (Waymouth medium, Life Technologies; Grand Island, NY) supplemented with 10 % fetal calf serum (Life Technologies), at 37 °C, 5 % CO₂ in a humidified incubator. They were routinely monitored for mycoplasma infection using fluorescence microscopy after DAPI staining to detect extranuclear nucleic acids.

Joy A., Kapoor M., Georges J., Butler L., Chang Y., Li C., Crouch A., Smirnov I., Nakada M., Hepler J., Marty M, & Feuerstein B.G. (2016). The role of AKT isoforms in glioblastoma: AKT3 delays tumor progression. Journal of neuro-oncology, 130(1), 43-52.

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Immunofluorescent Analysis of GSK-3 Phosphorylation

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Immunofluorescent analysis was performed as previously described (Huang et al., 2016 (link); Yu et al., 2018 (link); Hu et al., 2019 (link)). In brief, cryostat-coronal sections (20 μm) encompassing the entire midbrain were serially collected. Free-floating sections were pre-incubated in blocking solution containing 5% normal donkey serum and 0.3% Triton X-100 in 50 mM Tris-buffered saline (TBS, pH = 7.4) at room temperature for 1 h. Primary antibodies against p-GSK-3α Ser21 (Cell Signaling Technology, Cat. #9316, RRID:AB_659836), p-GSK-3β Ser9 (Cell Signaling Technology, Cat. #5558, RRID:AB_10013750), GSK-3α (Abcam, Cat. #ab40870, RRID:AB_732666), GSK-3β (Santa Cruz, Cat. #sc-9166, RRID:AB_647604), and TH (Merck Millipore, Cat. #AB9702, RRID:AB_570923) were dissolved in diluent and incubated with sections overnight at 4°C. After washing three times, sections were incubated with the secondary antibodies (Thermo Fisher or Jackson ImmunoResearch), which were conjugated with Alexa 488 or Alexa 555 at room temperature for 1 h. Finally, the sections were visualized under a confocal laser scanning microscope (LSM 780, Carl Zeiss, Germany).

Li J., Ma S., Chen J., Hu K., Li Y., Zhang Z., Su Z., Woodgett J.R., Li M, & Huang Q. (2020). GSK-3β Contributes to Parkinsonian Dopaminergic Neuron Death: Evidence From Conditional Knockout Mice and Tideglusib. Frontiers in Molecular Neuroscience, 13, 81.

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Protein Expression Analysis in Podocytes

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Antibodies were obtained from Cell Signaling Technology (pGSK3α (Ser21) #9316; pGSK3β(Ser9) #9323; pGSK3α/β(Ser21/9) #9331; p27Kip1 #2552; total β-catenin #8480; Cyclin B1 #12231; pcdk1(Tyr15)#4539; phosphoHistone3(ser10)#3377; cleaved Caspase 3 #9664; PARP #9542; YAP/TAZ #8418; pYAP (ser127) #4911; Ajuba #4897), Life Technologies (total GSK3 clone 21 A); Millipore (activated β-catenin clone 8E7), Sigma (β-actin; Podocin #0372; GAPDH), Acris (nephrin), Santa Cruz Biotechnology (Synaptopodin H-140; p27kip1), and Genetex (SV40 T antigen).

Hurcombe J.A., Hartley P., Lay A.C., Ni L., Bedford J.J., Leader J.P., Singh S., Murphy A., Scudamore C.L., Marquez E., Barrington A.F., Pinto V., Marchetti M., Wong L.F., Uney J., Saleem M.A., Mathieson P.W., Patel S., Walker R.J., Woodgett J.R., Quaggin S.E., Welsh G.I, & Coward R.J. (2019). Podocyte GSK3 is an evolutionarily conserved critical regulator of kidney function. Nature Communications, 10, 403.

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