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Phospho nf kb p65

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Phospho-NF-kB p65 is a primary antibody that specifically recognizes the phosphorylated form of the NF-kB p65 subunit. NF-kB p65 is a key transcription factor involved in the regulation of various cellular processes, including immune response, inflammation, and cell survival.

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Lab products found in correlation

Nf kb p65, by Cell Signaling Technology (5 mentions) Phospho p38 mapk, by Cell Signaling Technology (5 mentions) P38 mapk, by Cell Signaling Technology (3 mentions) Phospho p44 42 mapk, by Cell Signaling Technology (2 mentions) Phospho ikba, by Cell Signaling Technology (2 mentions) Phospho p38, by Cell Signaling Technology (2 mentions) Phospho jnk, by Cell Signaling Technology (2 mentions) Rpn2106, by GE Healthcare (1 mentions) Pgc1β, by Abcam (1 mentions) Phospho akt, by Cell Signaling Technology (1 mentions) β actin, by Merck Group (1 mentions) β actin antibody, by Cell Signaling Technology (1 mentions) Nitrocellulose membrane, by Cytiva (1 mentions) Appropriate imaging system, by LI COR (1 mentions) Nupage gel, by Thermo Fisher Scientific (1 mentions) P44 p42 mapk, by Cell Signaling Technology (1 mentions) Irak1, by Cell Signaling Technology (1 mentions) Supersignal west pico, by Thermo Fisher Scientific (1 mentions) Nf kb2 p100 p52, by Cell Signaling Technology (1 mentions) Gapdh cb1001, by Merck Group (1 mentions)

Close spelling variants of this product

Phospho-p65 (148 citations) NF-kB p65 (48 citations) NF-kB (47 citations) Anti-phospho-NF-kB p65 (9 citations) Phospho-NF-kB ( (4 citations) Phospho-NF-kB p65 Ser536 (4 citations) Anti-phospho- NF- kB p65 (Ser536) (3 citations) Rabbit anti-phospho-NF-kB p65 (2 citations)

15 protocols using phospho nf kb p65

1

Western Blot Analysis of Signaling Proteins

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Western blotting was performed following standard protocols. Primary antibodies reactive to phospho-NF-kB p65 (1:1000; Cell Signaling, 3033), NF-kB p65 (1:1000; Cell Signaling, 4764), phospho-AKT (1:500; Cell Signaling, 4060), AKT (1:1000; Cell Signaling, 4685), phospho-p38 MAPK (1:800; Cell Signaling, 4631), p38 MAPK (1:1000; Cell Signaling, 9212), PGC1β (1:1000; Abcam, 176328), and β-actin (1:20,000; Sigma, F3022) were used. Primary antibody binding was detected by enhanced chemoluminescence (GE Healthcare, RPN2106).
Rao S., Sigl V., Wimmer R.A., Novatchkova M., Jais A., Wagner G., Handschuh S., Uribesalgo I., Hagelkruys A., Kozieradzki I., Tortola L., Nitsch R., Cronin S.J., Orthofer M., Branstetter D., Canon J., Rossi J., D'Arcangelo M., Botling J., Micke P., Fleur L.L., Edlund K., Bergqvist M., Ekman S., Lendl T., Popper H., Takayanagi H., Kenner L., Hirsch F.R., Dougall W, & Penninger J.M. (2017). RANK rewires energy homeostasis in lung cancer cells and drives primary lung cancer. Genes & Development, 31(20), 2099-2112.
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2

Gingival Fibroblasts Activation by Saliva

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Human gingival fibroblasts were starved in serum-free medium overnight before being stimulated for 30 min with salivary pellet that had been washed four times. Whole human saliva, tumor necrosis factor alpha (TNF-α, 10 ng/mL) and Interleukin (IL)-1 (10 ng/mL) served as positive controls, and serum-free medium as the negative control. Cell extracts were separated by SDS-PAGE and transferred onto nitrocellulose membranes (Whatman, GE Healthcare, General Electric Company, Fairfield, CT). Primary antibody binding was accomplished with phospho-NF-kB p65 and β-actin antibodies (Cell Signaling Technology, Danvers, MA). Secondary antibody bindings were detected by near-infrared absorbing dyes with the appropriate imaging system (LI-COR Biosciences, Lincoln, NE).
Müller H.D., Cvikl B.B., Lussi A.A, & Gruber R.R. (2016). Salivary pellets induce a pro-inflammatory response involving the TLR4–NF-kB pathway in gingival fibroblasts. BMC Oral Health, 17, 15.
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3

Immunoblotting Protocol for Inflammatory Signaling

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Lysates were prepared in RIPA buffer (Cell Signaling Technologies) with 1mM PMSF per manufacturer's instructions. Proteins were separated on a NuPAGE gel (Invitrogen) and transferred to a PVDF membrane using the XCell II blotting system (Invitrogen). Membranes were blocked with 5% nonfat milk and incubated with primary antibody overnight at 4°C. Primary antibodies were purchased from Cell Signaling Technologies: IRAK1 (#4504), phospho-IkBa (#2859), IkBa (#4814), phospho-p38 MAPK (#4511), p38 MAPK (#8690), phospho-p44/42 MAPK (ERK1/2, #4370), p44/p42 MAPK (ERK1/2, #4695), NFkB2 p100/p52 (#4882), phospho-NFkB p65 (#3033), NFkB p65 (#8242) phospho-TBK1 (#5483), TBK1 (#3504). CXCL1 (PA1-29220, Thermo Fisher Scientific), GAPDH (CB1001), and total actin antibody (MAB1501) were purchased from EMD Millipore. HuR (sc-5261), IRAK1 (sc-5288), phospho-MAPKAPK2 (sc-293139), MAPKAPK2 (sc-393609), and TTP (sc-374305) were purchased from Santa Cruz Biotechnology. TRAF2 (592) was purchased from Medical & Biological Laboratories. Following washing, membranes were incubated with HRP-tagged anti-mouse IgG (1706516, Bio-Rad) or anti-rabbit IgG (NA934, GE Healthcare) and developed using SuperSignal West Pico or Femto substrate (Thermo Fisher Scientific).
Hornick E.E., Banoth B., Miller A.M., Zacharias Z.R., Jain N., Wilson M.E., Gibson-Corley K.N., Legge K.L., Bishop G.A., Sutterwala F.S, & Cassel S.L. (2017). Nlrp12 mediates adverse neutrophil recruitment during influenza virus infection. Journal of immunology (Baltimore, Md. : 1950), 200(3), 1188-1197.
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4

Quantifying Cellular NAMPT Signaling

Cited 1 time
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Reagents were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise noted. Antibodies for mTOR, Rictor, UCHL1, phospho-NF-kB p65, phospho-AktSer473 were purchased from Cell Signaling Technologies (Danvers, MA), β-actin and NF-kB from Invitrogen (Carlsbad, CA), goat anti-mouse IgG (Horseradish Peroxidase) from Jackson ImmunoResearch Laboratories. The anti-human goat NAMPT pAb was custom-generated as previously described10 (link),11 (link). The eNAMPT-neutralizing humanized mAb was provided by Aqualung Therapeutics (Tucson, AZ) as previously described11 (link). See Supplemental Materials and Methods for more details.
Bermudez T., Sammani S., Song J.H., Hernon V.R., Kempf C.L., Garcia A.N., Burt J., Hufford M., Camp S.M., Cress A.E., Desai A.A., Natarajan V., Jacobson J.R., Dudek S.M., Cancio L.C., Alvarez J., Rafikov R., Li Y., Zhang D.D., Casanova N.G., Bime C, & Garcia J.G. (2022). eNAMPT neutralization reduces preclinical ARDS severity via rectified NFkB and Akt/mTORC2 signaling. Scientific Reports, 12, 696.
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5

Cell Culture Materials and Reagents

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Cell culture materials were purchased from Irvine Scientific (Irvine, CA) or GIBCO (Grand Island, NY) except for skeletal muscle growth medium, which was obtained from Lonza (Walkersville, MD). [3H] 2-deoxyglucose, [14C]-L-glucose and [3H]-palmitate were obtained from Perkin Elmer (Boston, MA). All other chemicals were reagent grade and purchased from Sigma Chemical (St. Louis, MO), except for AG-1X8 ion exchange resin (Bio-Rad, Richmond, CA). Electrophoresis reagents were from Bio-Rad or Invitrogen (Carlsbad, CA). Primary antibodies were obtained from the following sources: IkBa (catalog #9242), phospho-p44/42 MAPK (#9106), p44/42 MAPK (#4695), phospho-p38 MAPK (#9216), p38 MAPK (#9212), phospho-NF-kB p65 (#13346), NF-kB p65 (#8242) (Cell Signaling Technology, Beverly, MA), phospho-JNK (#sc-6254), JNK (#sc-571), MyoD (#sc-760) (Santa Cruz Biotechnology, Santa Cruz, CA), b-actin (#NB600-503) (Novus, Littleton, CO). Fluorescently labeled secondary antibodies and blocking buffer were obtained from Licor (Lincoln, NE).
Ciaraldi T.P., Ryan A.J., Mudaliar S.R, & Henry R.R. (2016). Altered Myokine Secretion Is an Intrinsic Property of Skeletal Muscle in Type 2 Diabetes. PLoS ONE, 11(7), e0158209.
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6

Western Blot Analysis of NF-kB Signaling

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Total protein was separated using Radio-Immunoprecipitation Assay (RIPA) buffer (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Total protein extract was incubated with 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to polyvinylidene fluoride (PVDF) membranes and blocked with 5% skim milk at room temperature for 1 h. The membranes were incubated with primary antibodies against NF-kB p65, phospho-NF-kB p65 (1:1000; Cell Signaling Technology, Danvers, MA, USA) at 4 °C overnight, then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (Cell Signaling Technology) at room temperature for 2 h. Band intensities were standardized against β-tubulin and the relative density was analyzed on a Molecular Imager ChemiDoc XRS System (Bio-Rad Laboratories, Hercules, CA, USA) using enhanced chemiluminescence reagent (Thermo Fisher Scientific, Waltham, MA, USA).
Hu J., Wang D., Wu H., Yang Z., Yang N, & Dong J. (2019). Long non-coding RNA ANRIL-mediated inflammation response is involved in protective effect of rhein in uric acid nephropathy rats. Cell & Bioscience, 9, 11.
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7

Olive Leaf Extract Cytotoxicity Evaluation

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NB cell lines were seeded in 6-well plates, as already mentioned above. The day after seeding, cells were treated with 200 and 300 μM of OLE, as already described above. After 72 h of treatment, cells were collected and processed for cell signaling evaluation by FCM. Specifically, the expression of cleaved caspase 3, cleaved caspase 7, cleaved caspase 8, p53, cyclin D1, Bcl-2, phospho-Bcl-2, NF-kB p65, and phospho-NF-kB p65 was determined, according to the manufacturer’s instruction (Cell Signaling Technology Inc., Danvers, MA, USA).
Morandi F., Bensa V., Calarco E., Pastorino F., Perri P., Corrias M.V., Ponzoni M, & Brignole C. (2021). The Olive Leaves Extract Has Anti-Tumor Effects against Neuroblastoma through Inhibition of Cell Proliferation and Induction of Apoptosis. Nutrients, 13(7), 2178.
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8

Probing Cellular Signaling Pathways

Cited 1 time
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Primary antibodies used in this work were β-actin (Abcam, ab8226), SNAI1 EC3 (ref20 (link).), PY99 (Santa Cruz, sc7020), PDGFR-β (Cell Signaling, 3169), ERK1/2 (Cell Signaling, 9102), p-ERK1/2 (Cell Signaling, 9109), Akt (Cell Signaling, 4691), p-Akt (Cell Signaling, 4060), FAK (Cell Signaling, 3285), p-FAK (Cell Signaling, 3281), VE-Cadherin (Abcam, ab33168), VCAM-1 (Abcam, ab98954), MMP-9 (Abcam, ab38898), Snail (ref20 (link).), MT1-MMP (Abcam, ab51074), CD31/PCAM-1 (1:10, SC-506) Fibronectin (DAKO, A0245), Beta 1 integrin (Cell Signaling, 34971 S), Beta 3 integrin (Cell signaling, 13166 S), NF-KB p65 (Cell signaling, 8242 S), Phospho-NF-KBp65 (Cell signaling 3033 S) and Collagen I (Abcam, ab34710). Secondary antibodies used were anti-Mouse or anti-Rabbit IgG Antibody DyLight™ 680 or 800 Conjugate.
Other compounds used were PDGF-BB (Peprotech, 100-14B), BAPN: 3-Aminopropionitrile fumarate salt (Sigma, A3134), FAK inhibitor, PF-573228 (PZ0117, Sigma), ERK inhibitor, U0126-monoethanolate (Sigma, U120) and PI3K/Akt inhibitor LY294002 hydrochloride (Sigma, L9908).
Herrera A., Herrera M., Guerra-Perez N., Galindo-Pumariño C., Larriba M.J., García-Barberán V., Gil B., Giménez-Moyano S., Ferreiro-Monteagudo R., Veguillas P., Candia A., Peña R., Pinto J., García-Bermejo M.L., Muñoz A., García de Herreros A., Bonilla F., Carrato A, & Peña C. (2018). Endothelial cell activation on 3D-matrices derived from PDGF-BB-stimulated fibroblasts is mediated by Snail1. Oncogenesis, 7(9), 76.
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9

Evaluating M-CSF, RANKL, and TNF Signaling

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Human and murine M-CSF, sRANKL, and TNF were purchased from Peprotech (Rocky Hill, NJ, USA). Inhibitors, SP2509, RN-1, ORY-1001, and Torin1, were purchased from Cayman Chemical Company (Ann Arbor, MI). The antibodies used were as follows: LSD1, phospho-NFkB p65, Phospho-4E-BP1, HIF1a, E2F1, PHD-2 (Cell Signaling Technologies, Danvers, MA), p38, c-Myc (BioLegend, San Diego, California) and Swine Anti-Rabbit Immunoglobulins/HRP (DAKO, Hovedstaden, Denmark).
Doi K., Murata K., Ito S., Suzuki A., Terao C., Ishie S., Umemoto A., Murotani Y., Nishitani K., Yoshitomi H., Fujii T., Watanabe R., Hashimoto M., Murakami K., Tanaka M., Ito H., Park-Min K.H., Ivashkiv L.B., Morinobu A, & Matsuda S. (2022). LSD1 metabolically integrates osteoclast differentiation and inflammatory bone resorption through HIF-1α and E2F1. Arthritis & rheumatology (Hoboken, N.J.), 74(6), 948-960.
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10

PM10-Induced Signaling Pathway Analysis

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To determine the signaling pathways elicited by PM10, NPDFs were pretreated with the following signaling pathway inhibitors: SB203580 (p38 inhibitor, 10 μmol/L; Sigma-Aldrich), C-Jun (curcumin, 10 μmol/L; Sigma-Aldrich) or BAY117082 (NF-kb inhibitor, 2.5 μmol/L; Sigma-Aldrich). After a 1 h treatment of the inhibitors, the cells were incubated with PM10 (200 μg/mL) for 6 h and then lysed in LIPA buffer. Lysates were separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked with blocking solution and incubated with antibodies to NF-kb (Cell signaling Technology, Danvers, MA, USA), p38 (Cell signaling Technology), C-Jun (Cell signaling Technology), phospho-NF-kb p65 (Cell signaling Technology), phospho-p38 (Cell signaling Technology), phospho-C-Jun (Cell signaling Technology), and β- actin. Blots were visualized using horseradish peroxide-conjugated secondary antibodies and an Enhanced chemiluminescence (ECL) system (Pierce, Rockford, IL, USA).
Lee H.J, & Kim D.K. (2022). Effect of Airborne Particulate Matter on the Immunologic Characteristics of Chronic Rhinosinusitis with Nasal Polyps. International Journal of Molecular Sciences, 23(3), 1018.
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