TRAF3 Antibody 4729S from Cell Signaling Technology

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PLoS Pathogens
April 19 2012

Fig 1: Co-expression of LMP1/2A decreases B cell proliferation and TRAF2 levels compared to LMP1.(A–C) Purified splenic CD43- B cells were stimulated with combinations of IL-4, LPS, anti-IgM and anti-CD40. Proliferation as measured by 3[H]-thymidine incorporation is shown for 8 experiments with 8 mice per genotype. (D) Levels of TRAF2 and TRAF3 message were compared by RT-PCR using the ??CT method with HPRT as a housekeeping gene. Fold change in TRAF2 and TRAF3 expression in transgenic B cells compared to wildtype is shown for at least 4 experiments with 9 mice per genotype. (E) Immunoblot analysis shows TRAF2 (~53 kDa), TRAF3 (~62 kDa) and GAPDH (~38 kDa) expression in purified B cells from WT, LMP1, LMP2A and LMP1/2A mice and TRAF2-expressing CHO-K1 cells. Each lane represents a single mouse, and the data are representative of several immunoblot analyses with between 6 and 12 mice per genotype. (F) Quantification of TRAF2 signal normalized to GAPDH and wildtype TRAF2 signal is shown as an average of all mice from each genotype. For a description of quantification, see Methods. LMP1 bars have been placed first as LMP1 B cells gave the largest responses. Data are represented as mean ± standard error. *, P<0.05; **, P<0.01; ***, P<0.001 by one way ANOVA.
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Protein & cell
December 01 2020

Fig 2: TRIM35 catalyzes K48-linked polyubiquitination and degradation of IAV PB2. (A) Co-IP and IB analysis of HEK293T cells expressing TRIM35-V5 and WSN (H1N1) PB2. 10 µg/mL MG132 was added into cells at 6 h post-transfection (right panel). (B) Co-IP and IB analysis of HEK293T cells expressing TRAF3-Flag, TRIM35-V5, with or without WSN (H1N1) PB2. (C) IB analysis of HEK293T cells expressing WSN (H1N1) PB2 and increasing amounts of TRIM35-V5. (D) IB analysis of HEK293T cells expressing TRIM35-V5 or control vector for 36 h, followed by infection with WSN (H1N1) virus. (E) Co-IP and IB analysis to assess WSN (H1N1) PB2 ubiquitination in HEK293T cells expressing WSN (H1N1) PB2, with or without TRIM35-V5, along with HA-ubiquitin (K48) or HA-ubiquitin (K48R). (F) Co-IP and IB analysis to assess WSN (H1N1) PB2 ubiquitination in HEK293T cells expressing WSN (H1N1) PB2, with or without TRIM35-V5. (G) Minigenome assay in HEK293T cells expressing WSN (H1N1) PB2, PB1, PA, and NP, pHH21-SC09NS F-Luc, pRL-TK, along with control vector, TRIM35-V5, or both TRIM35-V5 and HA-ubiquitin. Results are expressed relative to Renilla luciferase activity. (H) Co-IP and IB analysis to assess WSN (H1N1) PB2 ubiquitination in HEK293T cells expressing WSN (H1N1) PB2, HA-ubiquitin (K48), along with V5-tagged TRIM35 or TRIM35 DelRING mutant. (I) IB analysis of HEK293T cells expressing V5-tagged TRIM35 DelRING mutant or control vector for 36 h, followed by infection with WSN (H1N1) virus. (J) Replication of WSN (H1N1) virus (MOI = 0.1) in HEK293 cells transfected for 24 h to express V5-tagged TRIM35 DelRING mutant or control vector, determined by plaque assay on MDCK cells. IB analysis was shown in the bottom panel. (K) Co-IP and IB analysis to assess K48-linked ubiquitination of WSN (H1N1) PB2 or its mutants in HEK293T cells expressing V5-tagged WSN (H1N1) PB2 or its mutants, GST-TRIM35, and HA-ubiquitin (K48). (L) IB analysis of HEK293T cells expressing WSN (H1N1) PB2 K736R mutant and increasing amounts of TRIM35-V5. Data are representative of at least three independent experiments. Means ± SD are shown in (G) and (J, upper panel) (n = 3). Two-tailed unpaired t-test was used for the statistical analysis, ***P < 0.001
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Protein & cell
December 01 2020

Fig 3: TRIM35 interacts with TRAF3. (A) Co-immunoprecipitation (co-IP) and IB analysis of HEK293T cells expressing TRIM35-V5 along with Flag-tagged RIG-I, VISA, TRAF3, TBK1, or IRF3. WCL, IB analysis of whole cell lysates without IP. (B) GST pull-down assay of purified His-tagged TRAF3 and lysates of HEK293T cells expressing GST-TRIM35. (C and D) Co-IP and IB analysis of the interaction between TRIM35 and TRAF3 in mouse peritoneal macrophages infected for 0–12 h with WSN (H1N1) virus (C) or SeV (D). (E) Binding of V5-tagged TRIM35 or its truncation mutants with TRAF3-Flag in transiently transfected HEK293T cells, as determined by co-IP and IB analysis. Human TRIM35 domains and truncation mutants were shown on top. (F) Binding of the V5-tagged TRIM35 PRY/SPRY domain with TRAF3-Flag in transiently transfected HEK293T cells, as determined by co-IP and IB analysis. (G) Binding of Flag-tagged TRAF3 or its truncation mutants with TRIM35-V5 in transiently transfected HEK293T cells, as determined by co-IP and IB analysis. Human TRAF3 domains and truncation mutants were shown on top. (H) Binding of the GST-tagged TRAF3 RING domain with TRIM35-V5 in transiently transfected HEK293T cells, as determined by co-IP and IB analysis. (I) Co-localization of TRIM35 and TRAF3 proteins in HEK293T cells expressing TRIM35-V5 and TRAF3-Flag for 36 h. Mitochondria and nuclei were stained using MitoTracker Red and DAPI, respectively. Data are representative of at least three independent experiments
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PLoS pathogens
May 01 2018

Fig 4: Both Smurf1 and the deubiquitinase activity of OTUD1 are required for OTUD1-mediated downregulation of MAVS/TRAF3/TRAF6 and inhibition of interferon antiviral response.(A-C) HEK293T cells were transfected with Flag-MAVS (A), or Flag-TRAF3 (B), or Flag-TRAF6 (C), together with or without shSmurf1 and Myc-OTUD1 as indicated. The whole cell lysates were analyzed by immunoblotting. (D-F) HEK293T cells were transfected with Flag-MAVS (D), or Flag-TRAF3 (E), or Flag-TRAF6 (F), together with or without shSmurf1 and Myc-OTUD1 as indicated. Immunoprecipitation and immunoblot analysis were carried out as indicated. (G) HEK293T cells were transfected with or without shSmurf1 and Myc-OTUD1 as indicated. After 48 hr, cells were infected with SeV for 12 hr. Relative mRNA expression of IFNß was determined by q-PCR. The data were shown as fold change normalized to that in the shCON group without Myc-OTUD1. (H) Immunoblot analysis of Smurf1, MAVS, TRAF3 and TRAF6 proteins in whole cell lysates of HEK293T cells transfected with empty vector, wild-type Flag-OTUD1, or catalytically inactive Flag-OTUD1 (CH). (I) Otud1-/- MEFs were transfected with empty vector, Flag-OTUD1, or Flag-OTUD1 (CH), and then infected with SeV for 12 hr. Relative IFNß mRNA expression was determined by q-PCR. The data were shown as fold change normalized to that in the uninfected control group. (J) HEK293T cells were transfected with empty vector, or Flag-OTUD1 wild type, or Flag-OTUD1 (CH), and then infected with VSV for 12 hr. Relative VSV viral RNA expression was analyzed by q-PCR. The data were shown as (I). *P<0.05 and **P<0.01, NS, not significant (P>0.05) (unpaired t-test). Error bars represent the mean and s.d., and all data are representative of three independent experiments.
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Cells
September 25 2022

Fig 5: Effect of non-viable Lactobacillus gasseri strains on signaling pathway in porcine alveolar macrophages after the activation of Toll-like receptor 3 (TLR3). Alveolar macrophages were stimulated with non-viable L. gasseri TMT36 (HK36), TMT39 (HK39), or TMT40 (HK40) and then challenged with the TLR3 agonist poly(I:C). The proteins from lysed cells were extracted at the indicated time points and separated by SDS-PAGE. Western-blot was performed to quantify TRAF3/ß-actin and p-IRF3/IRF3 ratios. Intensities of proteins bands were calculated from peak area of densitogram by using image Lab software. Non-lactobacilli treated alveolar macrophages challenged with poly(I:C) were used as controls. The results represent data from three independent experiments. Significant differences were observed when compared to polyI(:C)-challenged control porcine alveolar macrophages. * (p < 0.05), ** (p < 0.01).

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Cell Signaling Technology for
TRAF3 Antibody

The TRAF3 Antibody from Cell Signaling Technology is a Rabbit Polyclonal antibody to TRAF3. This antibody recognizes Homo Sapiens, Human, Mouse, Rat, and Non-Human Primate antigen. The TRAF3 Antibody has been validated for the following applications: Precipitation, Immunoprecipitation, and Western Blot.

Description

TRAF3 Antibody detects endogenous levels of total TRAF3 protein. Cross reactivity was not detected with other family members at physiological conditions

Product Specs
ItemTRAF3 Antibody
CompanyCell Signaling Technology
Price Pricing Info Supplier Page View Company Product Page
Catalog Number4729S
Quantity100 µl
ApplicationsW, IP
ReactivityH, M, R, Mk
TargetTRAF3
HostRabbit
NCBI Gene AliasesCAP-1, CAP1, CD40bp, CRAF1, IIAE5, LAP1, RNF118
NCBI Full Gene NameTNF receptor associated factor 3
ClonalityPolyclonal
Research AreaApoptosis
Molecular Weight62

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(1) White matter hyperintensities modify relationships between corticospinal tract damage and motor outcomes after strokemedRxivNovember 3, 2023Jennifer K. Ferris, Bethany P. Lo, Giuseppe Barisano, Amy Brodtmann, Cathrin M. Buetefisch, Adriana B. Conforto, Miranda H. Donnelly, Natalia Egorova-Brumley, Kathryn S. Hayward, Mohamed Salah Khlif, et al.4805, p = 0.748), stroke lesion volume (W = 5097, p = 0.717), or severity of motor impairment (W = 4729, p = 0.618) (see Supplementary Figure 2). In individuals with mild WMHs, motor impairment was significantly

(2) Effects of TRAF3 on the proliferation and migration of lung adenocarcinoma depend partly on pyroptosisBMC CancerOctober 5, 2023Wangjia Wang, Shiqi Wang, Min Wang, Yamei Ma, Wanting Hu, Binsha Wu, Chichi Li, Dan Zhangmin, the membranes were then incubated overnight with primary antibodies listed as follows: TRAF3 (#4729, Cell Signaling Technology, Danvers, MA, USA), IL-1β(#12,703, Cell Signaling Technology), cleaved IL-1β(#83,186,

(3) Influence of Race and High Laminar Shear Stress on TNFR1 Signaling in Endothelial CellsInternational Journal of Molecular SciencesSeptember 29, 2023Maitha Aldokhayyil, Dulce H. Gomez, Marc D. Cook, Andreas N. Kavazis, Michael D. Roberts, Thangiah Geetha, Michael D. Brownwere as follows: TNFR1 (cat. no. 3736), TACE (cat. no. 6978), TRAF2 (cat. no. 4724), TRAF3 (cat. no. 4729), and TRAF5 (cat. no. 41658) from Cell Signaling Technology (Beverly, MA, USA). β-actin (cat. no. 3700)

(4) Transcriptome Profiling of Vero E6 Cells during Original Parental or Cell-Attenuated Porcine Epidemic Diarrhea Virus InfectionVirusesJune 23, 2023Ouyang Peng, Yu Wu, Fangyu Hu, Yu Xia, Rui Geng, Yihui Huang, Siying Zeng, Guangli Hu, Chunyi Xue, Hao Zhang, et al.Australia, AB2838282), IRF3 (Proteintech, 11312-1-AP), TRAF3 (Cell Signaling Technology, Danvers, MA, USA, 4729S), PEDV-N, and GAPDH (Cell Signaling Technology) to detect respective proteins. Subsequently, the blots

(5) Macrophage fumarate hydratase restrains mtRNA-mediated interferon productionNatureMarch 1, 2023Hooftman A, Peace CG, Ryan DG, Day EA, Yang M, McGettrick AF, Yin M, Montano EN, Huo L, Toller-Kawahisa JE, et al.(2920), p-JNK (9255), JNK (9252), p-ERK1/2 (9101), ERK1/2 (4695), p-p38 (4511), p-38 (9212), TRAF3 (4729), p-p65 (3033) and GAPDH (2118) antibodies were purchased from Cell Signaling. Anti-goat IL-1β (AF-401-NA)

(6) The ULK3 kinase is a determinant of keratinocyte self-renewal and tumorigenesis targeting the arginine methylomeNature CommunicationsFebruary 16, 2023Sandro Goruppi, Andrea Clocchiatti, Giulia Bottoni, Emery Di Cicco, Min Ma, Beatrice Tassone, Victor Neel, Shadhmer Demehri, Christian Simon, G. Paolo DottoULK3 (Santa Cruz # 517373, 1:100 dilution), H3K9ac (Upstate #06-942, 1:100 dilution), H3K27ac (Abcam # 4729, 1:100 dilution), Pol II (Upstate #05-623, 1:100 dilution) or nonimmune antibodies (Cell Signaling,

(7) Neutrophils and macrophages drive TNF-induced lethality via TRIF/CD14-mediated responsesScience ImmunologyDecember 23, 2022Hayley I. Muendlein, Wilson M. Connolly, James Cameron, David Jetton, Zoie Magri, Irina Smirnova, Edouard Vannier, Xudong Li, Amanda J. Martinot, Rebecca Batorsky, et al.against p38 (9212), p-p38 (4511), ERK1/2 (4696), p-ERK1/2 (4370), IκBα (4814), RIPK1 (3493), TRAF3 (4729), TBK1 (3504), NEMO (2685), CASP8 (8592), CASP3 (9665), CASP7 (9492), CASP1 (24232), and GAPDH (2118)

(8) The Tumor Suppressor Protein TRAF3 Modulates GSK3 Activity and Susceptibility of B Lymphoma Cells to GSK3 InhibitionCancers (Basel)October 14, 2022Emma L. Hornick, Laura L. Stunz, Shakoora Sabree, Xiaosheng Wu, Thomas E. Witzig, Gail A. Bishop(Santa Cruz Biotechnology (Dallas, TX, USA) #sc-1828, 1:1000, or Cell Signaling Technologies (CST, Danvers, MA, USA) Ab #4729, 1:100) or GSK3β (CST, #12456). Lysates were incubated with IP Ab for 2 h at 4 °C, then Ab-bound targets

(9) TRAF3 enhances type I interferon receptor signaling in T cells by modulating the phosphatase PTPN22Science signalingSeptember 27, 2022Hornick EL, Wallis AM, Bishop GA.(Millipore Sigma #05-1154, 1:1000 dilution), TRAF3 (Santa Cruz Biotechnology #sc-1828, 1:1000, or CST #4729, 1:100), or IFNAR1 (abcam #ab45172, 1:500). Lysates were incubated with the immunoprecipitating antibody

(10) Modulation of Alveolar Macrophages by Postimmunobiotics: Impact on TLR3-Mediated Antiviral Respiratory ImmunityCellsSeptember 25, 2022Tomokiyo M, Tonetti FR, Yamamuro H, Shibata R, Fukuyama K, Gobbato N, Albarracin L, Rajoka MSR, Kober AKMH, Ikeda-Ohtsubo W, et al.antibody from Cell Signaling Technology (Beverly, MA, USA) and TRAF3 rabbit polyclonal antibody (Cat. #4729), respectively. A 1,000 times dilution of antibodies were used at 4 °C. After washing with TBS-T buffer

(11) Hypoxia induces HIF1α-dependent epigenetic vulnerability in triple negative breast cancer to confer immune effector dysfunction and resistance to anti-PD-1 immunotherapyNature communicationsJuly 15, 2022Ma S, Zhao Y, Lee WC, Ong LT, Lee PL, Jiang Z, Oguz G, Niu Z, Liu M, Goh JY, et al.H3K27me3 antibody (Cell Signaling Technology, CAT# 9733, dilution 1:1000), H3K27ac antibody (Abcam, CAT# 4729, dilution 1:1000), and GAPDH antibody (Cell Signaling Technology, CAT# 2118, dilution 1:1000). Co-Immunoprecipitation The

(12) ZBP1 promotes inflammatory responses downstream of TLR3/TLR4 via timely delivery of RIPK1 to TRIFProceedings of the National Academy of Sciences of the United States of AmericaJune 14, 2022Muendlein HI, Connolly WM, Magri Z, Jetton D, Smirnova I, Degterev A, Balachandran S, Poltorak A.CASP3 (9665), CASP7 (9492), RIPK1 (3493), pRIPK1 (Ser166) (31122), pRIPK1 (Ser321) (83613), TRAF3 (4729), TBK1 (3504), NEMO (2685), A20 (5630), p65 (8242), p-p65 (3033), and GAPDH (2118) were purchased from

(13) SMAD4 TGF-β–independent function preconditions naive CD8+ T cells to prevent severe chronic intestinal inflammationJournal of Clinical InvestigationApril 15, 2022Ramdane Igalouzene, Hector Hernandez-Vargas, Nicolas Benech, Alexandre Guyennon, David Bauché, Célia Barrachina, Emeric Dubois, Julien C. Marie, Saïdi M’Homa Soudjamanufacturer’s protocol. IPs were performed using anti-SMAD4 (Abcam, EP618Y), anti-H3K27ac (Abcam, 4729), and rabbit IgG (Cell Signaling Technology, 2729s). Primer sequences are provided in Table 2. In vitro

(14) FAM167A is a key molecule to induce BCR-ABL-independent TKI resistance in CML via noncanonical NF-κB signaling activationJournal of experimental & clinical cancer research : CRMarch 3, 2022Yang T, Sim KY, Ko GH, Ahn JS, Kim HJ, Park SG.from Santa Cruz Biotechnology; anti-NIK (4994), anti-p100/p52 (4882), anti-Myc (2276), anti-TRAF3 (4729), anti-CHIP (2080), and anti-c-cbl (2747) antibodies were purchased from Cell Signaling Technology;

(15) Epithelial phenotype restoring drugs suppress macular degeneration phenotypes in an iPSC modelNature communicationsDecember 15, 2021Sharma R, George A, Nimmagadda M, Ortolan D, Karla BS, Qureshy Z, Bose D, Dejene R, Liang G, Wan Q, et al.µg/mL; ab11867, Abcam), C3aR1 (~1 µg/mL; 126250, Abcam), RELB (~2 µg/mL; 4922, CST), TRAF3 (190 ng/mL; 4729, CST), VIMENTIN (~10 µg/mL; ab92547, Abcam), CLDN19 (~10 µg/mL; ab74374, Abcam), C5b-9 (5 µg/mL; M0777,

(16) TRAF3 in T Cells Restrains Negative Regulators of LAT to Promote TCR/CD28 SignalingJournal of immunology (Baltimore, Md. : 1950)July 1, 2021Arkee T, Hostager BS, Houtman JCD, Bishop GA.anti-PTP1B (cat #5311, used to detect PTP1B association with Brk in Fig 5H), and rabbit anti-TRAF3 (cat #4729) were purchased from Cell Signaling Technologies (Danvers, MA). Mouse anti-PTP1B (Ab38675) Ab, used

(17) E3 ligase Nedd4l promotes antiviral innate immunity by catalyzing K29-linked cysteine ubiquitination of TRAF3Nature communicationsFebruary 19, 2021Gao P, Ma X, Yuan M, Yi Y, Liu G, Wen M, Jiang W, Ji R, Zhu L, Tang Z, et al.(Ser32,2859), TBK1 (3504), IRF3 (4302), JNK (9252), NF-κB p65 (8242), IκBα (4814), Nedd4l (4013), TRAF3 (4729), c-Rel (12707), Ubiquitin (3936), K63-linkage-specific polyubiquitin (5621), K48-linkage-specific polyubiquitin

(18) Rv3722c Promotes Mycobacterium tuberculosis Survival in Macrophages by Interacting With TRAF3Frontiers in cellular and infection microbiologyJanuary 1, 2021Lei Y, Cao X, Xu W, Yang B, Xu Y, Zhou W, Dong S, Wu Q, Rahman K, Tyagi R, et al.Phospho-MAPK family antibody sampler kit (Cell Signaling Technology, 9910), TRAF3 (Cell Signaling Technology, 4729), NF-κB2 p100/p52 (Cell Signaling Technology, 4882), Alexa Fluor® 488 Goat anti-mouse IgG (Invitrogen,

(19) TRIM35 mediates protection against influenza infection by activating TRAF3 and degrading viral PB2Protein & cellDecember 1, 2020Sun N, Jiang L, Ye M, Wang Y, Wang G, Wan X, Zhao Y, Wen X, Liang L, Ma S, et al.(sc-6933), anti-TRIM35 (sc-100880), and anti-ubiquitin (sc-8017) were from Santa Cruz. Anti-TRAF3 (4729), anti-MAVS (24930), anti-K63-linkage specific polyubiquitin (5621), anti-K48-linkage specific polyubiquitin

(20) Remodeling of active endothelial enhancers is associated with aberrant gene-regulatory networks in pulmonary arterial hypertensionNature CommunicationsDecember 1, 2020Armando Reyes-Palomares et al.Clarified lysates from 20 million cells per sample were incubated with 1–5 μg of antibody (H3K27ac [Abcam #4729], H3K4me1 [Cell Signaling Technology #5326], and H3K4me3 [Cell Signaling Technology 39751]) coupled

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PLoS Pathogens

Protein & cell

Protein & cell

PLoS pathogens

Cells

TRAF3 Antibody from Cell Signaling Technology

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Cell Signaling Technology

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TRAF3 Antibody from Cell Signaling Technology

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