Nf κb p50

Manufactured by Abcam

Sourced in United Kingdom, United States, China

NF-κB p50 is a protein subunit of the NF-κB transcription factor complex. It plays a central role in the regulation of genes involved in immune and inflammatory responses.

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Lab products found in correlation

Nf κb p65, by Abcam (3 mentions) Pol 2, by Santa Cruz Biotechnology (1 mentions) β tubulin, by Abcam (1 mentions) C jun h79, by Santa Cruz Biotechnology (1 mentions) Snrp70, by Abcam (1 mentions) C fos h 125, by Santa Cruz Biotechnology (1 mentions) Pparγ, by Cell Signaling Technology (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Gapdh, by Abcam (1 mentions) Hif 1α, by Abcam (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions) Gsk 3β, by Abcam (1 mentions) Alexa 647, by Thermo Fisher Scientific (1 mentions) Tcs sp2 aobs, by Leica (1 mentions) Iba 1, by Abcam (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Immobilon p polyvinylidene difluoride membrane, by Merck Group (1 mentions) C myc, by Merck Group (1 mentions) Twist, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions)

6 protocols using nf κb p50

Flow Cytometry Analysis of Infected Cells

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Analysis of infected cells by flow cytometry and live cell sorting were performed as previously described
[5 (link)]. Of note, in all experiments, analysis was limited to live cells by FSC/SSC gating at the time of data acquisition. Unless otherwise stated, infected cells were analyzed four days post-infection. Jurkat E6-1 cells were stained and analyzed for CD69 as previously described
[51 (link)] except that antibodies were conjugated to PE-Cy7 and 1 μL of antibody was used per 1 × 10⁵ cells (BD Biosciences). Jurkat cells were stained with PE-Cy7-NFκB p65 (pS529) (BD Biosciences) and NFκB p50 (Abcam) with Pacific Blue conjugated secondary antibody (Life Technologies) as previously described
[52 (link)].

Dahabieh M.S., Ooms M., Brumme C., Taylor J., Harrigan P.R., Simon V, & Sadowski I. (2014). Direct non-productive HIV-1 infection in a T-cell line is driven by cellular activation state and NFκB. Retrovirology, 11, 17.

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Antibody Panel for Cell Signaling Analysis

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The antibodies used were as follows: NF-κB p65 (Abcam; cat. no. ab7970), Pol II (Santa Cruz; cat. no. SC-899 and SC-900, mixed in a 1:4 ratio), β-tubulin (Abcam; cat. no.ab6046), SNRP70 (Abcam; cat. no ab51266), c-Fos (H-125, Santa Cruz; cat. no. sc-7202), c-Jun (H-79, Santa Cruz; cat. no. sc-1694), and NF-κB p50 (Abcam; cat. no. ab7971).

Luo X., Chae M., Krishnakumar R., Danko C.G, & Kraus W.L. (2014). Dynamic reorganization of the AC16 cardiomyocyte transcriptome in response to TNFα signaling revealed by integrated genomic analyses. BMC Genomics, 15, 155.

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Protein Expression Analysis in Adipocytes

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Primary antibodies against β-Actin, AKT, phosphyorylated AKT (pAKT), GAPDH, glycogen synthase kinase-3 beta (GSK-3β), phosphorylated GSK-3β (pGSK-3β) Ser-9, HIF-1α, NF-κB p65, NF-κB p50, PAI-1 and phosphoenolpyruvate carboxykinase (PEPCK) were purchased from Abcam (Cambridge, UK); those against fatty acid synthase (FASN), fatty acid binding protein-4 (FABP4), pJNK and peroxisome proliferator-activated receptor gamma (PPARγ) were purchased from Cell Signaling (Danvers, MA, USA); those against JNK, c-Jun, sterol regulatory element-binding protein (SERBP1) and VEGFA were purchased from Santa Crutz; and that against diglyceride acyltransferase 2 (DGAT2) was purchased from Genetex. Secondary anti-rabbit antibody was purchased from Santa Cruz; anti-mouse antibody, from GeneTex; and anti-goat antibody, from Millipore (Temecula, CA, USA). Clarity^TM Western ECL Substrate was purchased from Bio-Rad; TRIzol Reagent, from Invitrogen; and 3-isobutyl-methylxanthine (IBMX), dexamethasone (Dex), and insulin, from Sigma.

Cheng Y.J., Liu C.C., Chu F.Y., Yang C.P., Hsiao C.W., Chuang C.W., Shiau M.Y., Lee H.T., Tsai J.N, & Chang Y.H. (2020). Oxygenated Water Inhibits Adipogenesis and Attenuates Hepatic Steatosis in High-Fat Diet-Induced Obese Mice. International Journal of Molecular Sciences, 21(15), 5493.

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Analyzing NF-κB Activation in Microglia

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NF-κB activation was analyzed in the paraffin-embedded sections from the ex vivo experiment as in Dadsetan et al. (26 (link)). For immunofluorescence, primary antibodies were NF-κB p50 (1:200, Abcam) and Iba1 (1:300, Abcam), followed by donkey anti-mouse Alexa 488 and donkey anti-rabbit Alexa 647 secondary antibodies (1:400, Invitrogen) and DAPI staining. Slides were observed under a confocal microscope (Leica TCS-SP2-AOBS) at 63x magnification and 8 fields per rat were acquired.
Nuclear and cytoplasmic intensity of p50 subunit was analyzed using ImageJ. Nuclei were outlined using the ROI manager function on DAPI blue channel and the selection was applied on the green channel (p50) to measure fluorescence. Mean intensity for each nucleus was measured. For cytoplasmic analysis of p50 subunit of NF-κB, green channels were used, and cytosol of each cell was manually outlined using a freehand selection of ImageJ and mean intensity recorded. Results are expressed as nuclear/cytoplasmic ratio of p50 subunit of NF-κB. Number of microglial cells (red signal) expressing NF-κB (green signal) was manually counted.

Izquierdo-Altarejos P., Martínez-García M, & Felipo V. (2022). Extracellular Vesicles From Hyperammonemic Rats Induce Neuroinflammation in Cerebellum of Normal Rats: Role of Increased TNFα Content. Frontiers in Immunology, 13, 921947.

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Western Blotting Analysis of EMT Markers

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The western blotting analysis was carried out using Immobilon-P polyvinylidene difluoride membranes (Millipore, 28820 Single Oak Drive, Temecula, California 92590, USA) as described in previous studies 42 (link),43 (link). The cell lysates were separated on sodium dodecyl sulfate-polyacrylamide gels and the western blotting analysis was carried out with following antibodies against CSMD1, Cyclin D1 (Santa Cruz Biotechnology), Snail (Abgent Inc, CA, USA), E-cadherin, Vimentin, Twist, Zeb1 (Cell Signaling Technology, MA, USA), iNOS, NFκB-p65, NFκB-p50 (Abcam, Cambridge, CB4 0FL, UK), and c-MYC (Sigma Chemical Co., MO, USA), respectively. β-actin (TaKaRa Co. Ltd, Kusachi, Japan) was used as an internal control. The density of band was measured using the ChemiDoc™ XRS⁺ System (1000 Alfred Nobel Drive, Hercules, California 94547, USA) equipping with Image-Pro Plus software and Epson color image scanner. The data was normalized to the β-actin.

Chen X.L., Hong L.L., Wang K.L., Liu X., Wang J.L., Lei L., Xu Z.Y., Cheng X.D, & Ling Z.Q. (2019). Deregulation of CSMD1 targeted by microRNA-10b drives gastric cancer progression through the NF-κB pathway. International Journal of Biological Sciences, 15(10), 2075-2086.

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Protein Expression Analysis of Liver Tissue

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Total proteins were extracted from liver tissue by RIPA lysis buffer (Ruian BioTechnology, Shanghai). The concentration of protein was measured using a BCA protein assay kit (Pierce Biotechnology, Rockford, USA) according to the manufacturer’s protocols. Forty mg protein was loaded and subjected to dodecyl sulphate-polyacrylamide gel (SDS-PAGE) electrophoresis, and then transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, Bedford, MA, USA). The membranes were immunoblotted with the indicated antibodies and imaged using an ECL detection system (Tanon, Shanghai, China). NLRP3 (1:500); αSMA (1:500); Collagen I (1:500); p-IκB (1:500); p-NF-κB p65 (1:500); IL-1β (1:500); NF-κB p65 (1:500) were purchased from Biosynthesis Biotechnology Inc. (Beijing, China); NF-κB p50 (1:500) and TLR4 (1:500) were purchased from Abcam (Cambridge, MA, USA), anti-IκB (1:1,000); anti-caspase 1 (1:1,000); β-actin (1:2,000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Dong Z., Zhuang Q., Ning M., Wu S., Lu L, & Wan X. (2020). Palmitic acid stimulates NLRP3 inflammasome activation through TLR4-NF-κB signal pathway in hepatic stellate cells. Annals of Translational Medicine, 8(5), 168.

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