The human antibodies used were anti-∆Np63α, anti-CB1, anti-cIAP2/BIRC3, and anti-ID1 (GeneTex, Irvine, CA, USA). Anti-AKT and anti-Phospho-AKT-Ser473 (Cell Signaling Technology, Danvers, MA, USA); anti-E-cadherin (BD; Baltimore, MD, USA); anti-β-catenin (Thermo Scientific, Waltham, MA, USA).
Anti akt and anti phospho akt ser473
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-AKT and anti-Phospho-AKT-Ser473 are primary antibodies that specifically detect AKT and phosphorylated AKT at serine 473, respectively. These antibodies are intended for use in various immunodetection 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 β catenin, by Thermo Fisher Scientific (2 mentions)
Anti cb1, by GeneTex (2 mentions)
Anti ciap2 birc3, by GeneTex (2 mentions)
Anti e cadherin, by BD (1 mentions)
Anti ki67, by Santa Cruz Biotechnology (1 mentions)
Anti p53, by Cell Signaling Technology (1 mentions)
Anti bcl 2, by ABclonal (1 mentions)
Hrp conjugated goat anti rabbit igg, by Merck Group (1 mentions)
Oddyssey fc imaging system, by LI COR (1 mentions)
Western bright tm sirius, by Advansta (1 mentions)
Goat anti mouse igg, by Merck Group (1 mentions)
Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Anti icam 1, by Santa Cruz Biotechnology (1 mentions)
Anti parp 1, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti phospho erk, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Pvdf filter, by Bio-Rad (1 mentions)
Prestained protein marker, by Thermo Fisher Scientific (1 mentions)
5 protocols using anti akt and anti phospho akt ser473
1
Immunohistochemical Analysis of Cell Signaling
2
Antibody-based Protein Analysis in Cells
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Specific antibodies used were: anti-ΔNp63α, anti-CB1 and anti-cIAP2/BIRC3 (GeneTex, Irvine, CA, USA); anti-AKT and anti-Phospho-AKT-Ser473 (Cell Signaling Technology, Danvers, MA, USA); anti-β-catenin (Thermo Scientific, Waltham, MA, USA); anti-P53 (Cell Signaling Technology, Danvers, MA, USA); anti-Bcl-2 (Abclonal, Woburn, MA, USA) and anti-Ki67 (Santa Cruz Biotechnology INC., Santa Cruz, CA, USA).
3
FTY720-Induced Signaling Pathway Analysis
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Following treatment with FTY720 in complete media, cells in 100-mm dishes were lysed in ice-cold RIPA lysis buffer containing protease inhibitors (cocktail set III; Calbiochem, Merck Millipore) and phosphatase inhibitors (Halt phosphatase inhibitor cocktail; Thermo Fisher Scientific, Waltham, USA). The protein lysates were collected and subjected to western blot analysis. The primary antibodies used were anti-Akt and anti-phospho-Akt (Ser473) (1:1000 dilution; Cell Signalling, USA), anti-ERK and anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (1:1000 dilution; Cell Signalling, USA), and secondary antibodies were HRP-conjugated goat anti-rabbit IgG or goat anti-mouse IgG (Sigma-Aldrich). The protein bands were detected with chemiluminescent reagent, Western Bright TM Sirius (Advansta, CA, USA) and viewed in an Oddyssey FC Imaging system (Licor Biosciences, Cambridge, UK).
4
Protein Extraction and Western Blot Analysis
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Cells were lysed in a buffer containing 10 mM Tris HCl, pH 7.5, NaCl 150 mM, and Triton 1% supplemented with Protease Inhibitor Cocktails (Sigma Aldrich, Saint Louis, MO, USA) and the protein content was measured by a colourimetric assay (BIO-RAD, Hercules, CA, USA). Protein cell lysates (20 μg) were separated by SDS-PAGE under denaturing conditions, transferred onto a PVDF filter (BIO-RAD, Hercules, CA, USA), blocked with 5% milk and probed with primary antibodies: anti-ERK2, anti-phospho-ERK, anti-ICAM1, anti-PARP1, anti-pan-cathepsin and anti-GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA), and anti-AKT and anti-phospho-AKT (Ser-473) (Cell Signalling Technologies, Danvers, MA, USA). After primary antibody incubation, the blots were washed and incubated with horseradish peroxidase-conjugated secondary antibodies (ThermoFisher Scientific; Waltham, MA, USA). The proteins recognized by Abs were detected by ECL (Amersham biosciences; Little Chalfont, UK). Pre-stained protein markers (ThermoFisher Scientific; Waltham, MA, USA) were used as molecular size standards. All experiments were performed in triplicate.
5
Western Blot Analysis of ANGPTL4 and AKT
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Cells were harvested in sodium dodecyl sulfate (SDS) sample buffer. Equal amounts of proteins were separated by using 10% SDS-PAGE system, transferred onto the polyvinylidene fluoride membrane (Millipore, USA). The membranes were blocked with 5% non-fat dried milk. Then, the membrane was blotted with the primary antibodies: anti-ANGPTL4 antibody (1:1000, Santa Cruz, USA), the polyclonal rabbit anti-HA antibody (1:1000, MBL, Japan), Anti-AKT and anti-phospho-AKT (Ser473) (1:1000, Cell Signaling Technology, Danvers, MA, USA), anti-GAPDH antibody (1:3000, ZSGB-BIO, China), diluted with 1% non-fat dried milk in TBST. After washed with TBST, the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibody (1:3000, ZSGB-BIO, China). The membranes were washed with TBST and scanned by ChemiDoc XRS+ chemiluminescence imaging system (Bio-Rad, USA). Each examination was tested in triplicate, and GAPDH was served as the internal reference.
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