Curcumin, CCK-8 kit, DMSO, and N-acetyl cysteine were purchased from Sigma Chemical Co (St. Louis, MO, United States) (Caspase-9, caspase-3,cleaved caspase-3,PARP,XIAP,p-Akt,Akt,GSK3,P-GSK3,FOXO1,P-FOXO1,GAPDH,cIap1,cIap2, Bcl-2, Bcl-xl, caspase 8, and cleaved caspase-8 antibodies were obtained from Cell Signaling Technologies (Beverly, MA, United States). Bax, p-H2AX, and cytochrome c antibodies and cisplatin were procured from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, United States). Annexin V fluorescein isothiocyanate (FITC), Propidium Iodide (PI), and p-H2AX (pS139) antibodies were purchased from BD Biosciences (San Jose, CA). z-VAD-FMK was obtained from Calbiochem (San Diego, CA, United States). CellROX Green was obtained from Invitrogen (MA, United States). Curcumin was dissolved in DMSO and further diluted in the cell culture media for the treatment of cells, so that the final concentration of DMSO in wells is 0.1% at the highest concentration of Curcumin used in the study. Viability assays showed that 0.1% DMSO is non-toxic to the cells (data not shown).
P foxo1
Manufactured by Cell Signaling Technology
Sourced in United States
P-FoxO1 is a phospho-specific antibody that detects FoxO1 protein phosphorylated at specific serine residues. FoxO1 is a transcription factor that plays a role in various cellular processes. The antibody can be used to detect and quantify the phosphorylation of FoxO1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Foxo1, by Cell Signaling Technology (34 mentions)
P akt, by Cell Signaling Technology (27 mentions)
Gapdh, by Cell Signaling Technology (7 mentions)
P erk, by Cell Signaling Technology (6 mentions)
P gsk3β, by Cell Signaling Technology (6 mentions)
Bcl 2, by Cell Signaling Technology (5 mentions)
P ampk, by Cell Signaling Technology (5 mentions)
β actin, by Merck Group (5 mentions)
P foxo3a, by Cell Signaling Technology (5 mentions)
P mtor, by Cell Signaling Technology (5 mentions)
P 4ebp1, by Cell Signaling Technology (4 mentions)
β actin, by Santa Cruz Biotechnology (4 mentions)
Pvdf membrane, by Merck Group (4 mentions)
Histone h3, by Cell Signaling Technology (3 mentions)
P stat3, by Cell Signaling Technology (3 mentions)
Nf κb, by Cell Signaling Technology (3 mentions)
Gapdh, by Bioworld Technology (3 mentions)
Ampkα, by Cell Signaling Technology (3 mentions)
α tubulin, by Merck Group (3 mentions)
Flag tag (flag), by Merck Group (3 mentions)
58 protocols using p foxo1
1
Curcumin-induced Apoptosis in Cancer Cells
2
Immunohistochemical Analysis of Gastric Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissue specimens from clinical GC samples and xenograft tumors derived from GC cells were fixed with 10% neutralbuffered formalin, and 4-μm paraffin sections were prepared. After rehydration, sections were stained with hematoxylin and eosin for histologic assessment, or were immunostained after antigen retrieval using a Bond-max automated immunostainer (Leica Microsystems, Newcastle upon Tyne, UK). The primary antibodies used were against FOXO1 (1:40, C29H4, Cell Signaling Technology), phospho-FOXO1Ser256 (pFOXO1; 1:60, Cell Signaling Technology), CD31 (1:100, M20, Santa Cruz Biotechnology), HIF-1α (1:50, provided by Dr. Jong-Wan Park at Seoul National University), VEGF (1:200, C1, Santa Cruz Biotechnology), and SIRT1 (1:100, H300, Santa Cruz Biotechnology). Antibody binding was detected with the Bond Polymer Refine Detection Kit (Leica Microsystems). All immunostained sections were lightly counterstained with Mayer’s hematoxylin. Throughout the above analysis, negative controls were prepared by omitting the primary antibody. The results of immunostaining were evaluated by two pathologists (Y.K. and J.-S.P.), who were blinded to the origin of the samples. For statistical analysis, the results of immunostaining for proteins were considered positive if immunoreactivity was seen in ≥ 10% (cytoplasmic pFOXO1 and nuclear SIRT1) or ≥ 5% (nuclear HIF-1α) of the neoplastic cells.
3
Antibody Sources for Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary antibodies against CRM1, GAPDH, actin, RanBP1, and p53 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and antibodies against cyclin D1, Cdc25B, p27, p21, Foxo1, p-Foxo1, Akt, p-Akt, p-Rb1, and histone H3 were purchased from Cell Signaling Technology (CST, Beverly, MA, USA).
4
Protein Lysate Extraction and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein lysate were extracted using RIPA lysis buffer (Thermo Fisher Scientific, China) contained protease and phosphatase inhibitors (1:100, Thermo Fisher Scientific, China). Subsequently, proteins were electrophoresed by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene fluoride membranes (PVDF, Millipore, Billerica, USA). After blocked with 5% bovine serum albumin for 1 hour, the membranes were incubated with following primary antibodies at 4℃ overnight: RPS9 (Proteintech, 1:1000); P- mTOR, mTOR , P-FAK, FAK, P-FoxO3a, FoxO3a, P-FoxO1, FoxO1, P-Stat3, Stat3, NF-κB, P-AMPK, AMPK, P-Smad2, Smad2, P-AKT, AKT, TGF-β, P-Erk and Erk (1:1000, Cell Signaling Technology, USA); β-actin (1:10000, Sigma-Aldrich, USA). Next day, after washed in PBST for three times, the membranes were incubated with HRP-conjugated anti-mouse or anti-rabbit antibodies (1:5000, Sigma-Aldrich, USA) at room temperature for 1 hour. Finally, the bands were visualized and analyzed by SuperSignal West Femto Maximun Sensitivity Substrate (Thermo Fisher Scientific, USA).
5
Protein Expression and Localization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues and cells were homogenized in Tissue Protein Extraction Reagent or Mammalian Protein Extraction Reagent (Thermo). Nuclear and cytoplasmic components were isolated using NE-PER nuclear and cytoplasmic extraction kit (Thermo). Homogenates (20 μg of total protein) were separated by SDS-PAGE and transferred to nitrocellulose membranes.
Antibodies were used against the following proteins: Sirt6, Sirt1, Ac-H3K9, FoxO1, p-FoxO1 (Cell Signaling, Beverly, MA, USA), Ac-lysine, Sirt2, Sirt3 (Abcam, Cambridge, UK), ubiquitin (Santa Cruz Biochemicals, Dallas, TX, USA), Sirt4, lamin B, GAPDH (Bioworld Technology, St Louis Park, MN, USA), Sirt5, Sirt7, Ac-FoxO1 (LifeSpan Biosciences, Seattle, WA, USA), and HSP90 (Enzo Life Sciences, Plymouth Meeting, PA, USA).
Antibodies were used against the following proteins: Sirt6, Sirt1, Ac-H3K9, FoxO1, p-FoxO1 (Cell Signaling, Beverly, MA, USA), Ac-lysine, Sirt2, Sirt3 (Abcam, Cambridge, UK), ubiquitin (Santa Cruz Biochemicals, Dallas, TX, USA), Sirt4, lamin B, GAPDH (Bioworld Technology, St Louis Park, MN, USA), Sirt5, Sirt7, Ac-FoxO1 (LifeSpan Biosciences, Seattle, WA, USA), and HSP90 (Enzo Life Sciences, Plymouth Meeting, PA, USA).
6
Immunoblotting Analysis of Key Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblotting was conducted according to our standard protocols, described previously [29 (link)]. The protein was extracted, quantified, and separated on SDS-PAGE gels, and electro-transferred to nitrocellulose membranes. The membranes were blocked in 3% BSA, and incubated with primary antibodies for FoxO1, p-FoxO1 (rabbit monoclonal, Cell Signaling Technology Inc., Danvers, MA, USA), PI3K (rabbit polyclonal, Cell Signaling), AKT, p-AKT, JNK, p-JNK, ERK, p-ERK, p38, p-p38 (rabbit monoclonal, Cell signaling) and β-actin (mouse monoclonal, Sigma-Aldrich, St Louis, MO). The blots were exposed to HRP-conjugated secondary rabbit IgG antibodies or mouse IgG antibodies, and analyzed by enhanced chemiluminescence (ECL) western blotting detection system (GE HealthCare Bio-Sciences, Piscataway, NJ, USA).
7
Podocyte Autophagy Regulation Mechanisms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Aldo, rapamycin (RP), chloroquine (CQ), 3-methyladenine (3-MA), tunicamycin (Tun), tauroursodeoxycholic acid (TAUDC), and anti-β-actin antibody were purchased from Sigma (St Louis, MO). Antibodies against LC3, Akt, p-Akt, mTOR, p- mTOR, S6K1, p-S6K1, 4EBP1, p-4EBP1, GRP78, GRP94, CHOP, FOXO1, p-FOXO1, Rab5, and Rab7 were purchased from Cell Signaling Technology (Beverly, MA). Anti-Podocin, anti-Nephrin, and anti-p62 antibodies were obtained from Abcam (Cambridge, MA). P300, Ac-FOXO1, and nestin antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).
8
Palmitate-Induced Cellular Stress Mechanisms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Palmitate was purchased from Calbiochem (San Diego, CA, USA). DCF-DA (2′,7′-dichlorofluorescin diacetate) was obtained from Molecular Probes (Eugene, OR, USA). Antibodies against β-actin, GRP78, CHOP, ATF6α, p-PERK, sXBP-1, p-JNK, Akt, Foxo1, GSK3β, IRS-1, IRS-2, HSP90, and CPR were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against p-AKT, p-Foxo-1, Foxo-1, p-AKT, p-GSK3β, p-Tyr, p-IR, and IR were acquired from Cell Signaling Technology (Beverly, MA, USA). Chlorozoxazone and p-nitrophenol were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).
9
AKT Signaling Pathway Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chlorogenic acid (purity >98.5%, determined by high performance liquid chromatography (HPLC) ()) was purchased from Aladdin (Beijing, China). Insulin was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Metformin hydrochloride (Met) was sourced from Adams Reagent, Ltd. (Shanghai, China). N3-tag (3-azido-7-hydroxy-2H-chromen-2-one) was supplied by WuXiAppTec (Beijing, China). The primary antibodies for glyceraldehyde 3-phosphate dehydrogenase (GAPDH), phosphorylated (p)-AKT, AKT, p-GSK3β, GSK3β, p-FOXO1, and FOXO1, and secondary antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Alexa Fluor® 594-conjugated goat anti-rabbit immunoglobulin G (IgG) was purchased from Abcam (Cambridge, UK). Chemiluminescent horseradish peroxidase (HRP) substrates were from Millipore Corporation (Burlington, MA, USA). SC79 and AKT inhibitor VIII were purchased from MCE (Monmouth Junction, NJ 08852, USA). PHT-427 and AT7867 were from Selleck (Shanghai, China). The pcDNA3-AKT-pleckstrin homology (PH)-GFP and pcDNA3-AKT-PH(R25C)-GFP plasmids were purchased from Addgene (Cambridge, MA, USA). All cell culture reagents were purchased from GibcoBRL Life Technologies (Grand Island, NY, USA). All other used chemicals were of analytical grade.
10
Microglial Activation and Inflammatory Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s modified Eagle’s medium (DMEM), 0.25% Trypsin-EDTA solution and fetal calf serum (FCS) were purchased from Gibco-BRL (Grand Island, NY, USA). Lithium chloride (LiCl) and LPS (Coli 0111:B4) were purchased from Sigma-Aldrich (St. Louis, MO, USA). LY294002, WST-8 dye, RIPA buffer and the BCA kit were purchased from Beyotime (Shanghai, China). The specific mouse anti-rat ED8 (anti-CD11b/CD18) monoclonal antibody (a marker for complement receptor 3 of activated microglia) was purchased from AbD Serotec (Raleigh, NC, USA). Fluoroshield mounting medium with 4,6-diamidino-2-phenylindole (DAPI) was purchased from Abcam (Hongkong, China). Rat IL-6 Immunoassay Kit and Rat TNF-α Immunoassay Kit were obtained from R&D Systems, Inc. (Minneapolis, MN, USA). Rabbit anti-TLR4 polyclonal antibody was purchased from Abcam (Hongkong, China). Specific rabbit monoclonal antibodies against p-PI3K, p-Akt, p-FoxO1, PI3K, Akt, FoxO1 and GAPDH, and secondary anti-rabbit and anti-mouse antibodies were all purchased from Cell Signaling (Boston, MA, USA). A FITC-conjugated goat anti-mouse IgG antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!