The following antibodies and reagents were purchased from the sources indicated: ERK 1/2 (R&D Cat# 9107S); pERK 1/2 (R&D Cat# 9101S); CDX2 (LSbio Cat# LS-B9317); NFĸB (Abcam cat # 16502), pNFĸB p65 (Cell Signaling Cat # 3031S); Claudin 1 (Invitrogen Cat# 51-9000); Claudin 2 (Invitrogen Cat# 32-5600); Claudin 3 (Invitrogen Cat# 341700); Claudin 7 (Invitrogen Cat# 374800); goat anti-rabbit-HRP (Santa Cruz Biotechnology Cat# sc-2004); goat anti-mouse-HRP (Santa Cruz Biotechnology Cat# sc-2005) and; actin (Sigma-Aldrich Cat# A1978). U0126 (Cat # 19-147) was from Sigma-Aldrich.
P nf bp65
Manufactured by Cell Signaling Technology
Sourced in United States
The P-NF-κBp65 product is a phospho-specific antibody that detects the phosphorylated form of the NF-κB p65 subunit. NF-κB p65 (also known as RelA) is a critical transcription factor involved in the regulation of various cellular processes, including inflammatory responses, immune function, and cell survival. The phosphorylation of NF-κB p65 is a key event in the activation of the NF-κB signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse hrp, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit hrp, by Santa Cruz Biotechnology (1 mentions)
Claudin 1, by Thermo Fisher Scientific (1 mentions)
U0126, by Merck Group (1 mentions)
Claudin 2, by Thermo Fisher Scientific (1 mentions)
Claudin 3, by Thermo Fisher Scientific (1 mentions)
Claudin 7, by Thermo Fisher Scientific (1 mentions)
P erk1 2, by R&D Systems (1 mentions)
Erk1 2, by R&D Systems (1 mentions)
Recombinant mouse m csf, by R&D Systems (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Cell culture medium, by Thermo Fisher Scientific (1 mentions)
Trap staining kit, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mapk family antibodysampler kit 9926t, by Cell Signaling Technology (1 mentions)
Phospho mapk family antibody sampler kit9910t, by Cell Signaling Technology (1 mentions)
Rankl, by R&D Systems (1 mentions)
Phosphatase inhibitor, by Solarbio (1 mentions)
Ripa lysate, by Solarbio (1 mentions)
5 protocols using p nf bp65
1
Protein Expression and Signaling Analysis
2
Osteoclastogenesis Assay Protocol
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Mon was purchased from Solarbio (Beijing, China). Fetal bovine serum (FBS) and cell culture medium were obtained from Gibco-BRL (Sydney, Australia). Recombinant mouse M-CSF and RANKL were purchased from R&D Systems (Minneapolis, MN, USA). TRAP staining kit was obtained from Sigma (St. Louis, MO, USA). TRIZOL reagent was bought from Invitrogen. PrimeScript TM RT reagent Kit was obtained from TaKaRa Biotechnology. RAW264.7 cells were purchased from Cell Bank (Chinese Academy of Sciences, Shanghai, China). The inhibitors of protease and phosphatase were purchased from Solarbio (Beijing, China). Antibodies for p-p38, p38, p-JNK, JNK, p-ERK, ERK, p-NF-ĸB p65, NF-ĸB p65, IĸBα and β-actin were obtained from Cell Signaling Technology (MAPK Family Antibody Sampler Kit #9926T, Phospho-MAPK Family Antibody Sampler Kit #9910T, and NF-κB Family Antibody Sampler Kit #9936).
3
Analysis of CPST Protein Expression Regulation
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All solvents used for the extraction and analysis of CPST in our study were of analytical grade and obtained from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Agilent XB C18 column was purchased from Agilent Technologies, USA. RIPA lysate, phosphatase inhibitors and PMSF were obtained from Beijing Solarbio Science Technology Co., Ltd. (Beijing, China). BCA protein concentration determination kit was purchased from Beyotime Biotechnology (Nanjing, China). Rabbit anti mouse myeloid differentiation primary response gene 88 (MyD88), NF-ĸBp65, p-NF-кBp65, NLRP3, apoptosis associated speck like protein containing CARD (ASC), caspase-1, cleaved caspase-1, IL-18, IL-1β, gasdermin-D (GSDMD) and uncoupling protein 1 (UCP1) were obtained from Cell Signaling Technology (MA, USA). Rabbit anti mouse thioredoxin-interacting protein (TXNIP) was purchased from Affinity Bioreagents (MA, USA). Rabbit anti-mouse TLR4 and β-actin were obtained from AB clonal Biotechnology (Wuhan, China). All primary antibody dilutions were 1:1000. WesternBright ECL prime detection reagent, SDS-PAGE gel kit, PVDF membrane were obtained from Boshide Biotechnology Company (Wuhan, China). Other regents used in the present study were of analytical grade.
4
Western Blot Analysis of Signaling Proteins
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p-NF-кBp65, p-JAK2, p-STAT3, NF-кBp65, JAK2, STAT3 antibody, and goat anti-rabbit IgG were from Cell Signaling Technology, China. The total protein was extracted from HUVECs according to the standard procedures. Protein samples (40 μg) were separated by 10% SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) and then transferred into PVDF membranes (Yiyuan Biotechnology, Guangzhou, China). The membrane was blocked with 5% nonfat dry milk solutions. After washing the PVDF membranes with TBST (tris buffered saline tween), the PVDF membranes were incubated overnight at 4°C with the above antibody, respectively. It was followed by secondary antibody for 2 h with goat anti-rabbit IgG. After washing, the membrane was developed with ECL kit (Yiyuan Biotechnology, Guangzhou, China) and detected with VILBER Fusion FX5 system (VILBER, Guangzhou, China).
5
Elacridar and DTT Protocol for Cellular Assays
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Elacridar and dithiothreitol (DTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), sterile phosphate-buffered saline (PBS), and penicillin/streptomycin antibiotics were obtained from Gibco (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Atlanta Biologicals (Flowery Branch, GA, USA). Total protein measurement reagents using the bicinchoninic acid (BCA) method were obtained from Pierce (Rockford, IL, USA). Antibodies used were mouse monoclonal antibody against light chain LRP1 (Abcam, Cambridge, MA, USA), mouse monoclonal antibody C-219 against P-gp from BioLegend (San Diego, CA, USA), mouse monoclonal antibody against BCRP (Cell signaling; Boston, MA, USA). Monoclonal antibodies for claudin-5, ZO-1, and HRP-labeled secondary antibodies were purchased from Invitrogen (Carlsbad, CA, USA), goat polyclonal antibodies against actin (C-11) and Matrix Metallopeptidase 9 (MMP9) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies for pre- and postsynaptic proteins SNAP-25 and PSD-95, respectively, were purchased from GeneTex (Irvine, CA, USA), antibodies for IκB-α, p-IκB-α, NF-кB p65, and p-NF-кB p65 were purchased from Cell Signaling. All other reagents and supplies were purchased from VWR (West Chester, PA, USA) and Fisher Scientific (Hampton, NH, USA).
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