Anti traf3

Manufactured by Santa Cruz Biotechnology

Anti-TRAF3 is a research antibody product developed by Santa Cruz Biotechnology. It is designed to detect and quantify TRAF3 protein expression in various samples. TRAF3 is a member of the TRAF family of proteins and plays a role in signal transduction pathways.

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Lab products found in correlation

Anti β actin, by Santa Cruz Biotechnology (2 mentions) Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (2 mentions) Anti actin, by Merck Group (1 mentions) Anti snrnp200, by Merck Group (1 mentions) Anti rela, by Santa Cruz Biotechnology (1 mentions) Anti nhp2l1, by Abcam (1 mentions) Anti ddx3x, by Fortis Life Sciences (1 mentions) Anti ddx60, by Abcam (1 mentions) Anti irf3, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated secondary antibody, by Bio-Rad (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Western lighting chemiluminescence reagent plus, by PerkinElmer (1 mentions) Anti pink1, by Abcam (1 mentions) Anti k63 ubiquitin, by Merck Group (1 mentions) Anti parkin, by Abcam (1 mentions) Anti gfp, by OriGene (1 mentions) Anti yap1, by Abcam (1 mentions) Anti flag, by OriGene (1 mentions) Anti myc, by OriGene (1 mentions) Anti ubiquitin k48, by Merck Group (1 mentions)

4 protocols using anti traf3

Comprehensive Cell Lysis and Immunoblotting

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Cells were washed twice with ice-cold phosphate-buffered saline (PBS; Wisent), harvested and lysed in 10mM Tris-HCl, 100mM NaCl, 0.5% Triton X-100, pH7.6 with EDTA-free Protease Inhibitor Cocktail (Roche). Cell lysates were clarified by centrifugation at 13,000 g for 20 min at 4°C and subjected to sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE). Western blot analysis was performed using mouse anti-PHF5A (Abnova), anti-IRF3 (Santa Cruz), anti-TRAF3 (Santa Cruz), anti-RIG-I (Alexis Biochemicals), anti-ACTIN (Chemicon International), anti-TBK1 (Imgenex and Santa Cruz), anti-IKBKE (Santa Cruz), anti-TUBULIN (ICN), anti-GAPDH (RDI) and rabbit anti-SNRNP200 (Sigma-Aldrich), anti-SF3A1 (Santa Cruz), anti-RELA (Santa Cruz), anti-NHP2L1 (Abcam), anti-DDX3X (Bethyl), anti-DDX60 (Abcam), TRIF (Cell Signaling), anti-ISG56 (Novus Biologicals), anti-MDA5 (Alexis Biochemicals), anti-MAVS (Alexis Biochemicals), anti-IKBKE (eBioscience), STAT1 (ABCAM), STAT1 tyr701 (ABCAM), IFNAR1 (Santa Cruz) and anti-IRF3-P-ser386 (Abcam). HRP-conjugated secondary antibodies were from Bio-Rad. The chemiluminescence reaction was performed using the Western Lighting Chemiluminescence Reagent Plus (PerkinElmer).

Tremblay N., Baril M., Chatel-Chaix L., Es-Saad S., Park A.Y., Koenekoop R.K, & Lamarre D. (2016). Spliceosome SNRNP200 Promotes Viral RNA Sensing and IRF3 Activation of Antiviral Response. PLoS Pathogens, 12(7), e1005772.

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Antibody Immunoblotting for Cellular Signaling

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Antibodies against phosphorylated and total IRF3, IKKε, TBK1, NF-κB p65, ERK, JNK, and p38 were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-TRAF3, anti-β-actin, and HRP-conjugated secondary antibodies were from Santa Cruz. Anti-Myc, anti-Flag, anti-HA, and anti-GFP were from Origene. Anti-PINK1, anti-Parkin, and anti-YAP1 were from Abcam Biotechnology (Cambridge, UK). Anti-K48-ubiquitin and anti-K63-ubiquitin were from Millipore (Kenilworth, USA).

Zhou J., Yang R., Zhang Z., Liu Q., Zhang Y., Wang Q, & Yuan H. (2019). Mitochondrial Protein PINK1 Positively Regulates RLR Signaling. Frontiers in Immunology, 10, 1069.

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Western Blot Analysis of Proteins

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Proteins were extracted from the ultracentrifugation pellets and separated on a denatured SDS-polyacrylamide gel before transfer to a polyvinylidene difluoride membrane (0.45 um; Millipore, Bedford, MA, USA). Afterwards, the membranes were blocked with 5% non-fat dry milk in Tris-buffered saline with Tween 20 (TBST) for 1 hour. The membranes were probed with primary anti-TRAF3 (1:200, Santa Cruz, sc-949), anti-PTEN (1:1000, Cell Signaling, #9559), anti-p65 (1:200, Santa Cruz, sc-372), anti-NIK (1:200, Santa Cruz, sc-7211) or anti-NFATc1 (1:1000, Cell Signaling, #8032), respectively. To check the amount of proteins transferred to nitrocellulose membrane, GAPDH was used as control and detected by an anti-GAPDH (1:500; Santa Cruz, sc-25778) antibody. The membranes were then washed three times with TBST and incubated for 1 hour with HRP-conjugated secondary antibodies (1:2000; Santa Cruz, sc-2004). The antigen-antibody complexes were visualized using SuperSignal^® West Dura Extended Duration Substrate (Thermo Scientific, Prod # 34075). Supplementary Fig. 7 depicts uncropped western blots.

Liu J., Li D., Dang L., Liang C., Guo B., Lu C., He X., Cheung H.Y., He B., Liu B., Li F., Lu J., Wang L., Shaikh A.B., Jiang F., Lu C., Peng S., Zhang Z., Zhang B.T., Pan X., Xiao L., Lu A, & Zhang G. (2017). Osteoclastic miR-214 targets TRAF3 to contribute to osteolytic bone metastasis of breast cancer. Scientific Reports, 7, 40487.

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Immunoblotting and Immunofluorescence Antibody Dilutions

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Antibodies used for immunoblotting and their corresponding dilutions were anti-MAVS (Santa Cruz Biotech sc-365334; 1:500), anti-TOM20 (Santa Cruz Biotech sc-11415; 1:300), anti-alpha Sr-1 (Abcam ab28052; 1:1,500), anti-β-actin (Santa Cruz Biotech sc-1615-hrp, 1:2,000), anti-MFN2 (Cell Signaling #9482; 1:800), anti-TBK1 (Cell Signaling #3504; 1:1,000), anti-phospho-TBK1 (Ser172) (Cell Signaling #5483; 1:700), goat anti-rabbit IgG-HRP (Millipore #12-348; 1:2000), and goat anti-mouse IgG-HRP (Millipore #12-349; 1:2,000). Primary antibodies used for immunofluorescence were anti-MAVS (Santa Cruz Biotech sc-365334; 1:100 dilution), anti-TOM20 (Abcam ab56783; 1:100 dilution), anti-FACL4 (Thermo PA5-27137; 1:50), anti-PMP70 (Abcam ab3424; 1:650), anti-MFN2 (Cell Signaling #9482; 1:50), anti-TRAF3 (Santa Cruz Biotech sc-1828or Thermo Fisher Scientific PA1-41107; 1:50), anti-alpha Sr-1 (Abcam ab28052; 1:50), anti-vimentin (Thermo Fisher Scientific PA1-16759; 1:2,000), anti-GM130 (BD Biosciences #610822; 1:250), anti-reovirus σ1 5C6 and G5 ((95 (link)); mix of 1:2,500 each; Fig. 6B), and rabbit anti-reovirus T3D antisera (B. Sherry, unpublished; 1:2,000; Fig S5). Secondary antibodies were Alexa^® 488-, Alexa^® 594- or Alexa^® 647-conjugated goat anti-mouse, anti-rabbit or anti-chicken IgG (Thermo Fisher Scientific; 1:750 - 1:1,000).

Rivera-Serrano E.E., DeAngelis N, & Sherry B. (2017). Spontaneous Activation of a MAVS-dependent Antiviral Signaling Pathway Determines High Basal Interferon-β Expression in Cardiac Myocytes. Journal of molecular and cellular cardiology, 111, 102-113.

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