Anti-FoxO3a Rabbit Monoclonal Antibody

RPL5 regulates colon cancer cell proliferation and migration through MAPK/ERK signaling pathway. A HCT116 and RKO cells were treated with RPL5-targeting siRNAs (siRPL5–1 and siRPL5–2) or negative control siRNA (siNC). After 48 h, cell lysates were harvested, and the protein samples were separated by SDS-PAGE. The levels of MAPK/ERK signaling pathway related proteins P-MEK1/2, MEK1/2, P-ERK, ERK, FOXO3 and c-Myc were detected by western blotting. The band intensities of P-MEK1/2, MEK1/2, P-ERK, ERK, FOXO3 and c-Myc proteins were quantified relative to β-Tubulin and normalized to the siNC sample. The blots were cut prior to hybridization with antibodies during blotting. B TBHQ (50 μM) was added to HCT116 and RKO cells after 6 h transfection of siRPL5, and the proteins of each group were extracted 48 h later. Changes of proteins related to MAPK/ERK signaling pathway were detected by western blotting. The blots were cut prior to hybridization with antibodies during blotting. C HCT116 and RKO cells were transfected with siRPL5 and then added TBHQ (20 μM) to detect the effect of TBHQ on the proliferation inhibition of colon cancer cells caused by siRPL5 at 24 h and 48 h. D HCT116 and RKO cells were transfected with siRPL5 and then added TBHQ (20uM) to detect the effect of TBHQ on the inhibition of colon cancer cell migration caused by siRPL5 at 24 h and 48 h. (E) TBHQ (50 μM) was added to HCT116 and RKO cells after transfection of siRPL5 for 6 h, and cell cycle-related proteins (CDK4 and CyclinD1) and migration-related proteins (MMP2 and MMP9) were detected by western blotting. The band intensities of CDK4, CyclinD1, MMP2 and MMP9 proteins were quantified relative to β-Tubulin and normalized to the siNC sample. The blots were cut prior to hybridization with antibodies during blotting. The data are representative of three independent experiments. (* P < 0.05, ** P < 0.01, *** P < 0.001)
Index in PubMed under a CC BY license. PMID: 36384455

All lanes use the Antibody at 1:1K dilution for 1 hour at room temperature.

Western blot analysis of FOXO3A expression in MCF-7 cell lysate.

Immunofluorescent analysis of Hela cells, using FoxO3a Antibody.

Western blot analysis of FoxO3a using anti-FoxO3a antibody (M00252).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: control group-normal mouse hippocampal tissue lysates,
Lane 2: hippocampal tissue from Alzheimer’s disease model mouse,
Lane 3: hippocampal tissue from Alzheimer’s disease model mouse treated with a self-developed drug.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-FoxO3a antigen affinity purified polyclonal antibody (Catalog # PA1079) at 1:2000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with ChemiDoc MP system. A specific band was detected for FoxO3a at approximately 80-100 kDa. The expected band size for FoxO3a is at 71.3 kDa.