Cells were seeded in a 6 cm dish at 4 × 104 cells/mL in 5 mL DMEM growth medium and incubated for 24 h at 37 °C, with 5% CO2 in a humidified atmosphere. Afterwards, cells were treated with different gelatin samples at a final concentration of 75 or 150 μg/mL for 6 h. The procedure for analyses of phosphorylated Akt and mTOR was performed according to the method described previously [48 (link)]. Three antibodies namely APC (allophycocyanin)-conjugated anti-Akt1 antibody (Thermo Fisher Scientific, Waltham, MA, USA), FITC (fluorescein isothiocyanate)-conjugated anti-phospho-Akt (Ser473) antibody (Thermo Fisher Scientific, Waltham, MA, USA), and PE (phycoerythrin)-conjugated anti-phospho-mTOR (Ser2448) antibody (Thermo Fisher Scientific, Waltham, MA, USA) were used in the current experiment.
Apc allophycocyanin conjugated anti akt1 antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The APC (allophycocyanin)-conjugated anti-Akt1 antibody is a fluorescent-labeled antibody that binds to the Akt1 protein. Akt1 is a serine/threonine-protein kinase that plays a key role in cellular processes such as glucose metabolism, apoptosis, cell proliferation, transcription, and cell migration.
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Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Dmem culture medium, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt dimethyl sulfoxide dmso, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fitc devd fmk, by Thermo Fisher Scientific (1 mentions)
Fitc conjugated anti brdu antibody, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (1 mentions)
2 protocols using apc allophycocyanin conjugated anti akt1 antibody
1
Gelatin Modulates Akt and mTOR Signaling
2
Extraction and Analysis of Brown Algae
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S. glaucescens, a type of brown algae, which was collected from Penghu, Taiwan, was washed with fresh water, dried, and then kept at 4 • C until use. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Trypsin/EDTA, fetal bovine serum (FBS), RPMI-1640 culture medium, DMEM culture medium, penicillin, and streptomycin were obtained from Gibco (Grand Island, NY, USA). Tetramethylrhodamine ethyl ester (TMRE) was purchased from Molecular Probes, Invitrogen Corp., (Carlsbad, CA, USA). Antibodies including FITC (fluorescein isothiocyanate)-labelled anti-cytochrome c antibody, FITC-LEHD-FMK, FITC-DEVD-FMK, FITC-conjugated anti-BrdU antibody, FITC-conjugated anti-phospho-Akt (Ser473) antibody, PE-conjugated anti-phospho-mTOR (Ser2448) antibody, PerCP-eFluor 710-conjugated anti-phospho-S6 (Ser235, Ser236) antibody, and APC (allophycocyanin)-conjugated anti-Akt1 antibody were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Annexin V-FITC and PI were purchased from BD PharMingen (San Diego, CA, USA).
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