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Traf2

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TRAF2 is a protein that plays a role in the regulation of cellular signaling pathways. It functions as an adapter protein, facilitating the interaction between various proteins involved in signal transduction. TRAF2 is known to be involved in the activation of the NF-κB and MAPK signaling cascades.

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Lab products found in correlation

β actin, by Santa Cruz Biotechnology (3 mentions) Ire1α, by Santa Cruz Biotechnology (2 mentions) Bcl2 associated x (bax), by Santa Cruz Biotechnology (2 mentions) Sirna transfection reagent, by Santa Cruz Biotechnology (2 mentions) C rel, by Santa Cruz Biotechnology (2 mentions) Traf6, by Santa Cruz Biotechnology (2 mentions) Polyclonal antibodies generated against dream, by Santa Cruz Biotechnology (2 mentions) Nf κb2, by Santa Cruz Biotechnology (2 mentions) Tak1 rabbit mab d94d7, by Cell Signaling Technology (2 mentions) Phospho sapk jnk thr183 tyr185 rabbit mab 81e11, by Cell Signaling Technology (2 mentions) Sapk jnk rabbit mab 56g8, by Cell Signaling Technology (2 mentions) Ikkα pab, by Cell Signaling Technology (2 mentions) Phospho ikkα β ser176 180 pab, by Cell Signaling Technology (2 mentions) Ikkβ pab, by Cell Signaling Technology (2 mentions) Rip2 pab, by Cell Signaling Technology (2 mentions) P65 rela pab, by Merck Group (2 mentions) Lipids dimethyldioctadecylammonium bromide and chlolesterol for liposome preparation, by Merck Group (2 mentions) Nf κb1, by Santa Cruz Biotechnology (2 mentions) Anti β actin mab clone ac 15, by Merck Group (2 mentions) Control sirna, by Qiagen (2 mentions)

9 protocols using traf2

1

Bufalin-Induced Apoptosis in Leukemia Cells

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Cell lines K562 and K562/A02 cells were purchased from the Shanghai Institute of Cell Sciences, Chinese Academy of Sciences (Shanghai, PRC). ADM and bufalin were purchased from Sigma-Aldrich (St. Louis, MO). Nrf2, HO-1, P-gp, Bax, Bcl-2, cyctc, PAPR, IRE1α, TRAF2, JNK, p-JNK, and Caspase-12 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). The apoptosis detection kit was purchased from KeyGen Biotech (Nanjing, PR China). RT-PCR and HiPerfect transfection kits were purchased from Qiagen (Hilden, Germany).
Xie Y., Yan X, & Sun L. (2019). The Mechanism of Bufalin-Induced Apoptosis of K562/A02. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 2542-2552.
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2

Immunoprecipitation and Luciferase Assays

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ELISA kits were from eBioscience (ThermoFisher). Antibodies used for western blots and coimmunoprecipitation were: Arid5a, FLAG (Sigma Aldrich), β actin-HRP, α Tubulin-HRP (Abcam, Cambridge UK), IκBξ, Myc (Cell Signaling, Beverly MA), C/EBPβ (BioLegend, San Diego CA), IκBα, MCPIP1, TRAF2, Act1, YY1 (Santa Cruz Biotechnology, Santa Cruz CA). Protein G or Protein A beads (Roche) were used to pull-down antibodies during co-immunoprecipitation. Luciferase assays used the Dual-Luciferase Reporter Assay System (Promega) (6 (link)).
Amatya N., Childs E.E., Cruz J.A., Aggor F.E., Garg A.V., Berman A.J., Gudjonsson J.E., Atasoy U, & Gaffen S.L. (2018). IL-17 integrates multiple self-reinforcing, feed-forward mechanisms through the RNA-binding protein Arid5a. Science signaling, 11(551), eaat4617.
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3

Apoptosis Signaling Pathway Proteins

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The following antibodies were used: caspase-1 (Adipogen), caspase-8 (Enzo; Cell Signaling), caspase-3 (Cell Signaling), RIPK1 (BD Biosciences), IL-1β (Genetex), TRAF2 (Santa Cruz Biotechnology), and β-actin (Santa Cruz Biotechnology).
Van Hauwermeiren F., Van Opdenbosch N., Van Gorp H., de Vasconcelos N., van Loo G., Vandenabeele P., Kanneganti T.D, & Lamkanfi M. (2022). Bacillus anthracis induces NLRP3 inflammasome activation and caspase-8–mediated apoptosis of macrophages to promote lethal anthrax. Proceedings of the National Academy of Sciences of the United States of America, 119(2), e2116415119.
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4

Western Blot Analysis of Signaling Proteins

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Western blot analysis was conducted according to the method of Yao et al. [38 (link)]. The supernatant was collected after lysed cells were centrifuged at 12,000 rpm at 4 °C for 15 min. Protein extracts were separated with SDS-PAGE and transferred to PVDF membrane. After blocking, p-p53, p-p38, p-ASK1, TRAF2, and β-actin antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were added and incubated. Membranes were washed and incubated in PBST and HRP-conjugated secondary antibody, respectively. The signals were detected using the Super Signal ELISA Pico Chemiluminescent Substrate (Thermo Fisher Scientific, Waltham, MA, USA).
Ren G., Shi Z., Teng C, & Yao Y. (2018). Antiproliferative Activity of Combined Biochanin A and Ginsenoside Rh2 on MDA-MB-231 and MCF-7 Human Breast Cancer Cells. Molecules, 23(11), 2908.
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5

Antibodies and Reagents for NF-κB Signaling

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Polyclonal antibodies generated against DREAM, ICAM-1, USF1, TRAF2, TRAF6, NF-κB1, NF-κB2, RelB, c-Rel, and IκBα were obtained from Santa Cruz Biotechnology, Inc. Anti-A20 mouse monoclonal antibody (mAb; 59A426) was from Calbiochem. c-Fos polyclonal antibody (pAb), phospho-TAK1 (Thr184/187) pAb, TAK1 rabbit mAb (D94D7), phospho-p38 MAPK (Thr/Tyr182) mAb (28B10), p38 MAPK pAb, phospho-SAPK/JNK (Thr183/Tyr185) rabbit mAb (81E11), SAPK/JNK rabbit mAb (56G8), IKKα pAb, phospho-IKKα/β (Ser176/180) pAb, IKKβ pAb, phospho-IκBα (Ser32) rabbit mAb (14D4), RIP1 pAb, and RIP2 pAb were from Cell Signaling. IKKγ mAb (clone 72C627) and DREAM mAb (clone 40A5) were obtained from Upstate. p65/RelA pAb was from Chemicon. Control siRNA was from Qiagen and hUSF1-siRNA (SMARTpool, cat # L003617) was from Dharmacon. siRNA transfection reagent was obtained from Santa Cruz Biotechnology, Inc. Lipids (dimethyldioctadecylammonium bromide and chlolesterol) for liposome preparation and anti-β-actin mAb (clone AC-15) were purchased from Sigma. PCR primers were custom-synthesized from Integrated DNA Technologies.
Tiruppathi C., Soni D., Wang D.M., Xue J., Singh V., Thippegowda P.B., Cheppudira B.P., Mishra R.K., DebRoy A., Qian Z., Bachmaier K., Zhao Y., Christman J.W., Vogel S.M., Ma A, & Malik A.B. (2014). The transcription factor DREAM represses A20 and mediates inflammation. Nature immunology, 15(3), 239-247.
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6

Western Blot Analysis of Cellular Fractions

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Equal amount of whole cell lysates, nuclear fraction, cytoplasmic fraction, and membrane fraction from cells of different treatment groups were used for Western blotting as we previously described [26 (link)]. The primary antibodies were against Sam68 (1:1000, Santa Cruz, sc-333), P-IKKαβ (1:100, Cell Signaling, 9958s), IKKβ (1:100, Cell Signaling, 2684s), P-IkBα (1:100, Cell Signaling, 2859S), p-NFκB p65 Antibody (Ser 276) (1:1000, Santa Cruz, sc-101749), Lamin A (1:1000, Santa Cruz, sc-20680), FiLamin A (1:1000, Abcam, ab3261), TRAF2 (1: 250, Santa Cruz, sc-876) , IL-1β (1:1000, Cell Signaling, 31202S), IL-6 (1:1000, Cell Signaling, 12912T),NF-κB p65 (1:1000, Cell Signaling, 8242T) and β-Actin (1:1000, Abcam, ab8227).
Han S., Xu S., Zhou J., Qiao A., Boriboun C., Ma W., Li H., Biyashev D., Yang L., Zhang E., Liu Q., Jiang S., Zhao T.C., Krishnamurthy P., Zhang C., Richard S., Qiu H., Zhang J, & Qin G. (2019). Sam68 impedes the recovery of arterial injury by augmenting inflammatory response. Journal of molecular and cellular cardiology, 137, 82-92.
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7

Western Blotting of Key Signaling Proteins

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The following primary antibodies were used for Western blotting: anti-ALK (Life Technology, Monza, IT; mouse 1:2000), TRAF1 (Santa Cruz, rabbit 1:1000) and TRAF2 (Santa Cruz, Heidelberg DE; mouse 1:1000) anti-Actin Millipore (Merk-Millipore, Milano IT; mouse 1:2000), and anti MYC (rabbit 1:1000), antiphospho-STAT3 (rabbit 1:1000), anti STAT3-Y705 (rabbit 1:1000), anti SHP2 (rabbit 1:1000), anti SHP2-Y542 (rabbit 1:1000), anti ERK1/2-Thr202/Tyr204 (rabbit 1:1000), p50 NFkB (rabbit 1:1000), and p52NFkB (rabbit 1:1000) from Cell Signaling Technology (Danvers, MA). Western blotting and immunoiprecipaitation studies were performed as previously described {Zamo, 2002 (link) #5422;Piva, 2006 (link) #1201;Boccalatte, 2009 (link) #639}.
Abate F., Todaro M., van der Krogt J.A., Boi M., Landra I., Machiorlatti R., Tabbo’ F., Messana K., Barreca A., Novero D., Gaudiano M., Aliberti S., Di Giacomo F., Tousseyn T., Lasorsa E., Crescenzo R., Bessone L., Ficarra E., Acquaviva A., Rinaldi A., Ponzoni M., Longo D.L., Aime S., Cheng M., Ruggeri B., Piccaluga P.P., Pileri S., Tiacci E., Falini B., Pera-Gresely B., Cerchietti L., Iqbal J., Chan W.C., Shultz L.D., Kwee I., Piva R., Wlodarska I., Rabadan R., Bertoni F, & Inghirami G. (2014). A novel Patient Derived Tumorgraft model with TRAF1-ALK Anaplastic Large Cell Lymphoma translocation. Leukemia, 29(6), 1390-1401.
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8

Antibody-Based Analysis of Neuronal Stress

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Mouse monoclonal antibodies against tyrosine phospho-Hydroxylase (Ser31), GAPDH, β-actin, 4-Hydroxy-2-nonenal (4-HNE), Nitro-tyrosine, CHOP, Fas, Bax, NFkB p65, Histone H1, TRAF2, IRE1α, and phospho-IKB-β were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rabbit monoclonal antibodies against phospho-p38 MAPK (Thr180/Tyr182) and phospho-JNK1/2 (Thr183/Tyr185) were purchased from Cell Signaling Technology (Beverly, MA, USA). The TdT-mediated dUTP Nick End Labeling (TUNEL) kits were purchased from Roche (Germany). MPP+ (1-methyl-4-phenylpyridinium), SDS, NP-40, while sodium deoxycholate, protease inhibitor cocktail was purchased from Sigma (St. Louis, MO, USA).
Kuo H.C., Lu C.C., Shen C.H., Tung S.Y., Hsieh M.C., Lee K.C., Lee L.Y., Chen C.C., Teng C.C., Huang W.S., Chen T.C, & Lee K.F. (2016). Hericium erinaceus mycelium and its isolated erinacine A protection from MPTP-induced neurotoxicity through the ER stress, triggering an apoptosis cascade. Journal of Translational Medicine, 14, 78.
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9

Antibodies and Reagents for NF-κB Signaling

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Polyclonal antibodies generated against DREAM, ICAM-1, USF1, TRAF2, TRAF6, NF-κB1, NF-κB2, RelB, c-Rel, and IκBα were obtained from Santa Cruz Biotechnology, Inc. Anti-A20 mouse monoclonal antibody (mAb; 59A426) was from Calbiochem. c-Fos polyclonal antibody (pAb), phospho-TAK1 (Thr184/187) pAb, TAK1 rabbit mAb (D94D7), phospho-p38 MAPK (Thr/Tyr182) mAb (28B10), p38 MAPK pAb, phospho-SAPK/JNK (Thr183/Tyr185) rabbit mAb (81E11), SAPK/JNK rabbit mAb (56G8), IKKα pAb, phospho-IKKα/β (Ser176/180) pAb, IKKβ pAb, phospho-IκBα (Ser32) rabbit mAb (14D4), RIP1 pAb, and RIP2 pAb were from Cell Signaling. IKKγ mAb (clone 72C627) and DREAM mAb (clone 40A5) were obtained from Upstate. p65/RelA pAb was from Chemicon. Control siRNA was from Qiagen and hUSF1-siRNA (SMARTpool, cat # L003617) was from Dharmacon. siRNA transfection reagent was obtained from Santa Cruz Biotechnology, Inc. Lipids (dimethyldioctadecylammonium bromide and chlolesterol) for liposome preparation and anti-β-actin mAb (clone AC-15) were purchased from Sigma. PCR primers were custom-synthesized from Integrated DNA Technologies.
Tiruppathi C., Soni D., Wang D.M., Xue J., Singh V., Thippegowda P.B., Cheppudira B.P., Mishra R.K., DebRoy A., Qian Z., Bachmaier K., Zhao Y., Christman J.W., Vogel S.M., Ma A, & Malik A.B. (2014). The transcription factor DREAM represses A20 and mediates inflammation. Nature immunology, 15(3), 239-247.
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