Human recombinant EGF and human recombinant TNF were purchased from Peprotech (Rocky Hill, NJ, USA). Gefitinib was kindly provided by AstraZeneca (Tokyo, Japan). TAPI-1 was from Calbiochem (San Diego, CA, USA). The following antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA): Total EGFR antibody (#4267), phospho-EGFR (Y1068) antibody (#3777), total AKT antibody (#9272), phospho-AKT (S473) antibody (#9271), total ERK1/2 antibody (#9102), phospho-ERK1/2 antibody (#4370), phospho-MKK3/6 antibody (#9236), total p38 MAPK antibody (#9212), phospho-p38 MAPK antibody (#9211), TACE antibody (#3976), and β-actin antibody (#4970). Meanwhile, antibodies specific to total MKK 3/6 (sc-13069) was purchased from Santa Cruz Biotechnology, Dallas, TX, USA. Cetuximab was purchased from Showa University Hospital (Shinagawa, Tokyo, Japan). Materials used for western blot analysis were from Bio-Rad Laboratories (Richmond, CA, USA). Dulbecco’s modified Eagle medium (DMEM) and all other materials were purchased from Sigma Chemical (St. Louis, MO, USA) unless otherwise stated. All pharmacological inhibitors and agonists were dissolved in DMSO and added to the medium with a final DMSO concentration of <0.1%.
Phospho akt s473 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Phospho-AKT (S473) antibody is a tool used to detect and quantify the phosphorylation of the AKT protein at the serine 473 residue. This antibody can be used in various analytical techniques, such as Western blotting, to study the activation and regulation of the AKT signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit igg, by Cell Signaling Technology (2 mentions)
β actin antibody, by Cell Signaling Technology (1 mentions)
Total akt antibody, by Cell Signaling Technology (1 mentions)
Phospho erk1 2 antibody, by Cell Signaling Technology (1 mentions)
Recombinant human egf, by Thermo Fisher Scientific (1 mentions)
Recombinant human tnf, by Thermo Fisher Scientific (1 mentions)
Total egfr antibody, by Cell Signaling Technology (1 mentions)
Total erk1 2 antibody, by Cell Signaling Technology (1 mentions)
Total p38 mapk antibody, by Cell Signaling Technology (1 mentions)
Phospho p38 mapk antibody, by Cell Signaling Technology (1 mentions)
Dulbecco s modified eagle medium dmem, by Merck Group (1 mentions)
Gefitinib, by AstraZeneca (1 mentions)
Et1608 64, by Huabio (1 mentions)
Et1602 47, by Huabio (1 mentions)
Akt antibody, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions)
Gotaq qpcr master mix, by Promega (1 mentions)
4 protocols using phospho akt s473 antibody
1
Investigating EGFR and Cell Signaling
2
CB-Induced Signaling Pathway Analysis
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Cells were treated with CB (0, 0.25, 0.5 μM) for 48 h and then the cell lysates were analyzed by immunoblotting. The primary antibodies used in this study were as follows: ATM antibody (ET1606-20, Huabio), phospho-ATM-S1981 antibody (ET1705-50, Huabio), phospho-AKT-S473 antibody (4060S, Cell Signaling Technology), AKT antibody (4685S, Cell Signaling Technology), phospho-mTOR-S2448 antibody (5536T, Cell Signaling Technology), mTOR antibody (2983T, Cell Signaling Technology), phospho-p70S6K-T389 antibody (9234S, Cell Signaling Technology), p70S6K antibody (9202S, Cell Signaling Technology), Bcl-2 antibody (ET1702-53, Huabio), active+pro Caspase-3 antibody (ET1608-64, Huabio), active Caspase-3 antibody (ET1602-47, Huabio), p53 antibody (ab26, Abcam), γH2AX antibody (201082-2A9, ZEN-BIOSCIENCE), phospho-p53-S15 antibody (YP0205, ImmunoWay), CHK2 antibody (ET1610-52, Huabio), phospho-CHK2-T68 antibody (29012-1-AP, Proteintech) and GAPDH antibody (60004-1-lg, Proteintech). The relative protein intensity was calculated by ImageJ software.
3
Phospho-Akt (S473) Western Blot
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Cells were plated at 200,000 per well in a 6-well plate and incubated with the compound for 4 hours. Lysates were made using RIPA buffer, run using sodium dodecyl sulfate polyacrylamide gel electrophoresis, transferred to polyvinylidene difluoride membrane and probed with phospho-Akt (S473) antibody (Cell Signaling) followed by antirabbit IgG (Cell Signaling). Band intensity was calculated using ImageJ ver. 1.42 (National Institutes of Health, Bethesda, MD).
4
Titanium Surface Analysis for Cell Behavior
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Two different titanium surfaces (discs) were investigated in this study, distinguished by the surface properties, as follows: Machined (Wo/DAE), and Dual Acid-Etched (W/DAE). The metallic TiO2-based alloys were generously donated by the S.I.N. (São Paulo, SP, Brazil). The cell culture flasks were purchased from TPP (Trasadingen, Switzerland), and the cell culture reagents from Nutricell (Campinas, SP, Brazil). Ripa buffer (R0278), Phosphatase inhibitor cocktail 2 (P5726), and bovine serum albumin (A7906) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Gotaq qPCR master mix (A6002) was purchased from PROMEGA (Madison, WI, USA). The following antibodies were purchased from Cell Signaling (Danvers, MA, USA): Akt Antibody (4691P, 60 kDa); PhosphoAkt (S473) Antibody (4060P, 60 kDa); Anti-Rabbit IgG (7074).
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