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C rel

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C-Rel is a laboratory equipment product manufactured by Cell Signaling Technology. It is a nuclear transcription factor that plays a crucial role in the regulation of gene expression. C-Rel is a member of the NF-κB family of proteins and is involved in various cellular processes, including immune response, inflammation, and cell growth and survival.

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Lab products found in correlation

Nf κb2, by Cell Signaling Technology (3 mentions) Cyclin d1, by Thermo Fisher Scientific (2 mentions) Hsp60, by Santa Cruz Biotechnology (2 mentions) Met clone c 28, by Cell Signaling Technology (2 mentions) Phospho erk p44 42, by Santa Cruz Biotechnology (2 mentions) Nfkbie, by Santa Cruz Biotechnology (2 mentions) On targetplus human nfkbie sirna smartpool, by Horizon Discovery (2 mentions) Subcellular protein fractionationkit for cell culture, by Thermo Fisher Scientific (2 mentions) Rela p65, by Cell Signaling Technology (2 mentions) Mab3211, by Merck Group (2 mentions) Bay 11 7082, by Cayman Chemical (2 mentions) Nuak1, by Cell Signaling Technology (2 mentions) Antibody against tubulin t5168, by Merck Group (2 mentions) Rela s536 p, by Cell Signaling Technology (2 mentions) A2066, by Merck Group (2 mentions) Hrp conjugated antibodies against mouse igg na931, by Cytiva (2 mentions) Rabbit igg na934, by Cytiva (2 mentions) Ros glo h2o2 assay, by Promega (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Stat3, by Cell Signaling Technology (2 mentions)

18 protocols using c rel

1

Protein Expression Analysis Protocol

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AR (D6F11; Cell Signaling), p62 (H00008878-M01; Abnova), SOD2 (AM7579a; Abgent), β-actin (sc-69879; Santa Cruz), RELA (6956S; Cell Signaling), NFKB1 (3035S; Cell Signaling), PSA (5365S; Cell Signaling), or c-REL (4727T; Cell Signaling).
Thomas-Jardin S.E., Dahl H., Kanchwala M.S., Ha F., Jacob J., Soundharrajan R., Bautista M., Nawas A.F., Robichaux D., Mistry R., Anunobi V., Xing C, & Delk N.A. (2019). RELA is sufficient to mediate interleukin-1 repression of androgen receptor expression and activity in an LNCaP disease progression model. The Prostate, 80(2), 133-145.
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2

Extensive Immunohistochemistry and Western Blot Analyses

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Immunohistochemistry was performed using the following antibodies: p16 (Ventana, catalog number 9517), p53 (Dako, catalog number GA6166), Cyclin D1 (ThermoFisher, catalog number RM-914-S), CDK4 (Invitrogen, catalog number AHZ0202), EGFR (Ventana, catalog number 790-4347), MDM2 (Invitrogen, catalog number 182403), Rb (BD Biosciences, catalog number 554136), Met (clone c-28), YAP1 (Cell Signaling, catalog number 4912), phospho-ERK p44/42 (clone D13,14.4E).
Antibodies for Western Blots were as follows: NFKBIE (Santa Cruz, catalog number sc-7155); RELA/p65 (Cell Signaling, 8242); p50/p105 (Cell Signaling, p/n 3035); cREL (Cell Signaling, p/n 4727); PARP (Cell Signaling, 9542); HSP60 (Santa Cruz, sc-1722). NFKBIE siRNAs were from Dharmacon (ON-TARGETplus Human NFKBIE siRNA SMARTpool). Cell fractionation was performed using the Subcellular Protein FractionationKit for Cell Culture (Thermo Scientific 78840). The M257 and M375 cell lines were provided by Dr. Antoni Ribas.
Shain A.H., Garrido M., Botton T., Talevich E., Yeh I., Sanborn J.Z., Chung J., Wang N.J., Kakavand H., Mann G.J., Thompson J.F., Wiesner T., Roy R., Olshen A.B., Gagnon A., Gray J.W., Huh N., Hur J.S., Busam K.J., Scolyer R.A., Cho R.J., Murali R, & Bastian B.C. (2015). Exome sequencing of desmoplastic melanoma identifies recurrent NFKBIE promoter mutations and diverse activating mutations in the MAPK pathway. Nature genetics, 47(10), 1194-1199.
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3

NF-κB Pathway Antibodies and Assays

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Antibodies against p-RelA-S536 (#3033; 1:1000), RelA (#8242; 1:1000), RelB (#4922, 1:2000; IHC: #10544), c-Rel (#4727; 1:1000), NF-κB2 (#3017; 1:1000), LKB1 (#3050; 1:1000), and NUAK1 (#4458; 1:1000) were purchased from Cell Signalling Technology (Danvers, MA). Antibodies against lamin A/C (MAB3211; 1:500) and actin (A2066; 1:20000) was purchased from Millipore (Temecula, CA, USA). Antibody against tubulin (T5168; 1:20000) was purchased from Sigma. HRP-conjugated antibodies against mouse IgG (NA931; 1:10000) and rabbit IgG (NA934; 1:10000) were purchased from Cytiva. All antibodies were diluted in tris-buffered saline-Tween 20 containing 5% bovine serum albumin, with the exception of NUAK1 (tris-buffered saline-Tween 20 containing 5% nonfat milk). BAY 11-7082 was purchased from Cayman Chemical (cat. 10010266; Ann Arbor, MI, USA). ROS-Glo H2O2 Assay was purchased from Promega (cat. G8820).
Buensuceso A., Fritz J.L., Collins O., Valdés Y.R., Borrelli M.J., DiMattia G.E, & Shepherd T.G. (2022). Loss of LKB1-NUAK1 signalling enhances NF-κB activity in a spheroid model of high-grade serous ovarian cancer. Scientific Reports, 12, 3011.
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4

Extensive Immunohistochemistry and Western Blot Analyses

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Immunohistochemistry was performed using the following antibodies: p16 (Ventana, catalog number 9517), p53 (Dako, catalog number GA6166), Cyclin D1 (ThermoFisher, catalog number RM-914-S), CDK4 (Invitrogen, catalog number AHZ0202), EGFR (Ventana, catalog number 790-4347), MDM2 (Invitrogen, catalog number 182403), Rb (BD Biosciences, catalog number 554136), Met (clone c-28), YAP1 (Cell Signaling, catalog number 4912), phospho-ERK p44/42 (clone D13,14.4E).
Antibodies for Western Blots were as follows: NFKBIE (Santa Cruz, catalog number sc-7155); RELA/p65 (Cell Signaling, 8242); p50/p105 (Cell Signaling, p/n 3035); cREL (Cell Signaling, p/n 4727); PARP (Cell Signaling, 9542); HSP60 (Santa Cruz, sc-1722). NFKBIE siRNAs were from Dharmacon (ON-TARGETplus Human NFKBIE siRNA SMARTpool). Cell fractionation was performed using the Subcellular Protein FractionationKit for Cell Culture (Thermo Scientific 78840). The M257 and M375 cell lines were provided by Dr. Antoni Ribas.
Shain A.H., Garrido M., Botton T., Talevich E., Yeh I., Sanborn J.Z., Chung J., Wang N.J., Kakavand H., Mann G.J., Thompson J.F., Wiesner T., Roy R., Olshen A.B., Gagnon A., Gray J.W., Huh N., Hur J.S., Busam K.J., Scolyer R.A., Cho R.J., Murali R, & Bastian B.C. (2015). Exome sequencing of desmoplastic melanoma identifies recurrent NFKBIE promoter mutations and diverse activating mutations in the MAPK pathway. Nature genetics, 47(10), 1194-1199.
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5

TRAF1, NF-κB, and Apoptosis Signaling

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Cell Signaling Technology (CST) TRAF1 (#4715, Rabbit mAb), p105/50 (#3035, Rabbit mAb), RelA (#8242, Rabbit mAb), phospho-RelA (Ser536) (3033, Rabbit mAb), RelB (#4922, Rabbit mAb), cRel (#4727, Rabbit mAb), IκBα (#9247, Mouse mAb), IRF4 (#4964, Rabbit mAb), TAK1 (#4505, Rabbit mAb), V5 (#13202, Rabbit mAb), HRP-linked anti-mouse IgG(7076), FLIP (#8510, Rabbit mAb), HRP-linked anti-rabbit IgG (#7074) were used in this study at 1:1000 dilution. p100/52 (EMD Millipore #05–361, Mouse mAb, 1:1000), GAPDH (EMD Millipore #MAB374, Mouse mAb, 1:500) was used. S12 mouse monoclonal antibody against LMP1 was purified from hybridoma supernatant (104 (link)). The IKKβ inhibitor IKK-2 inhibitor VIII (ApexBio, #A3485), puromycin dihydrochloride (Thermo Fisher #A1113803), and hygromycin B (Millipore #400052).
Mitra B., Beri N.R., Guo R., Burton E.M., Murray-Nerger L.A, & Gewurz B.E. (2023). Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation. bioRxiv.
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6

RNA Expression and Protein Analysis of Th Cell and DC Markers

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qPCR was performed as previously described10 (link). Total RNA was extracted from cells by using an RNeasy Mini kit (Qiagen) according to the manufacturer's instructions. The mRNA levels of Dectin1, Il9, Ifng, Il4, Il5, Il13, Il17, Tnfsf15, Ox40l, Spi1, Irf4, Tbx21, Gata3 and Rorc by Th cells or DCs were analysed. Expression was normalized to the expression of the house-keeping gene Gapdh. Primer sets used for these analyses are listed in Supplementary Table 1.
Western blot assay was performed as previously described10 (link). Anti-mouse phosphorylated (p)-Syk, Syk, pRaf1, Raf1, p-IKKα/β, IκB-α, p65, p50, c-Rel, RelB, p52, β-actin and HDAC1 antibodies were purchased from Cell Signaling Technology (CST). RIPA Buffer (cat #: 9806) were purchased from CST. Nuclear extraction kit (cat #: 78833) was purchased from Thermo Scientific. Images have been cropped for presentation. Full-size images are presented in Supplementary Figs 14 and 15.
Zhao Y., Chu X., Chen J., Wang Y., Gao S., Jiang Y., Zhu X., Tan G., Zhao W., Yi H., Xu H., Ma X., Lu Y., Yi Q, & Wang S. (2016). Dectin-1-activated dendritic cells trigger potent antitumour immunity through the induction of Th9 cells. Nature Communications, 7, 12368.
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7

Profiling Signaling Pathways in Cell Lines

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All human cell lines were obtained from the American Type Culture Collection (ATCC), which performs authentication on its own cell lines. All the cell lines were cultured in DMEM supplemented with 10% heat-inactivated FBS at 37°C and 5% CO2 in a humidified incubator. Antibodies against total PDCD4 (CAT#:9535S), AKT1 (CAT#:75692), AKT2 (CAT#:3063), AKT3 (CAT#:14982), P65 (CAT#:8242), P50 (CAT#:3035), c-Rel (CAT#:4727), RelB (CAT#:10544), Tubulin (CAT#:2148s), Histone3 (CAT#:4499), Actin (CAT#:3700), STAT3 (CAT#:9139), ERK(CAT#:4695), FAK1 (CAT#:3285), phospho-STAT3 (CAT#:9145), phospho-ERK (CAT#:4370), and phospho-FAK1 (CAT#:8556) were purchased from Cell Signaling Technology. Antibody against VEGF (CAT#:ab52917) was purchased from Abcam. Human Cytokine Antibody Array C3 was purchased from Raybio.
Pin G., Huanting L., Chengzhan Z., Xinjuan K., Yugong F., Wei L., Shifang L., Zhaojian L., Kun H., Weicheng Y., Yingying L., Yongming Q, & Yanan Y. (2020). Down-Regulation of PDCD4 Promotes Proliferation, Angiogenesis and Tumorigenesis in Glioma Cells. Frontiers in Cell and Developmental Biology, 8, 593685.
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8

Western Blot Analysis of Signaling Proteins

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Total cell lysates and nuclear and cytoplasmic extracts were prepared from sorted and/or treated CD11c+ and CD11c− cells as described in text using lysis buffer (Cell signaling) and nuclear extraction kit (Panomics), respectively. Lysates (15 µg) were separated on 10% SDS-PAGE gels and then transferred onto PVDF membranes (Millipore). Nonspecific binding was blocked by incubation in blocking buffer (5% nonfat milk in TBST), followed by sequential incubations with the primary antibody and appropriate horseradish peroxidase-conjugated secondary antibody (Thermo Scientific), each diluted in blocking buffer. Immunoreactive protein was detected using enhanced chemiluminescence (ECL) reagents (Thermo Scientific). Antibodies against κBα (Santa Cruz biotech), phospho-IκBα, NF-κB subunit p65, c-rel, LSD1 (Cell Signaling), phospho IKKα/β, IKKα/β (Cell Signaling), MnSOD (EMD Millipore), Cu-ZnSOD (Enzo lifesciences) and β-actin (Cell signaling) were used for western blot analysis. Densitometric analysis was performed using the ImageJ software.
Khare A., Raundhal M., Chakraborty K., Das S., Corey C., Kamga C.K., Quesnelle K., St. Croix C., Watkins S.C., Morse C., Oriss T.B., Huff R., Hannum R., Ray P., Shiva S, & Ray A. (2016). Mitochondrial H2O2 In Lung Antigen Presenting Cells Blocks NF-κB Activation To Prevent Unwarranted Immune Activation. Cell reports, 15(8), 1700-1714.
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9

Western Blot Analysis of Signaling Proteins

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Cells were maintained in complete medium before the transfection assays, which are performed in medium without serum for 48 h and then lysed in RIPA buffer containing a mixture of protease inhibitors. Equal amounts of protein extract were resolved on SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, Italy), probed overnight at 4 °C with antibodies against: A20 (A-12, sc-166,692) and β-Actin (AC-15, sc-69,879) (Santa Cruz Biotechnology, DBA, Italy), CTGF (Origene, DBA, Milan, Italy), c-Rel (Cell Signaling Technology, Milan, Italy) and then revealed using the ECL™ Western Blotting Analysis System (GE Healthcare, Italy).
Santolla M.F., Lappano R., Cirillo F., Rigiracciolo D.C., Sebastiani A., Abonante S., Tassone P., Tagliaferri P., Di Martino M.T., Maggiolini M, & Vivacqua A. (2018). miR-221 stimulates breast cancer cells and cancer-associated fibroblasts (CAFs) through selective interference with the A20/c-Rel/CTGF signaling. Journal of Experimental & Clinical Cancer Research : CR, 37, 94.
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10

Protein Expression Analysis Protocol

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Primary antibodies directed against the following antigens were used: p50 (Abcam, Cambridge, USA), p65 (Cell Signaling Technology, Danvers, USA), c-Rel (Cell Signaling Technology), RelB (Santa Cruz Biotechnology, Dallas, USA) and p52 (Cell Signaling Technology), αSMA (Abcam), PAI-1 (Santa Cruz Biotechnology), pSmad3 (Abcam), GAPDH (Cell Signaling Technology), focal adhesion kinase, FAK (Abcam), SM22α (ThermoFisher Scientific, Waltham, USA), talin (Abcam), vinculin (SigmaAldrich, St. Louis, USA), actin (Merck Millipore, Burlington, USA), N-cadherin (Abcam) and Calnexin (ENZO Life Sciences, Belgium).
Micus L.C., Trautschold-Krause F.S., Jelit A.L., Schön M.P, & Lorenz V.N. (2021). NF-кB c-Rel modulates pre-fibrotic changes in human fibroblasts. Archives of Dermatological Research, 314(10), 943-951.
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