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Tnfaip3

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TNFAIP3 is a protein that plays a role in the regulation of inflammation and immune responses. It functions as a negative regulator of the NF-kappa-B signaling pathway, which is involved in the expression of genes related to inflammation, immunity, and cell survival.

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Lab products found in correlation

Stmn1, by Abcam (1 mentions) Proliferating cell nuclear antigen (pcna), by Abcam (1 mentions) Ccnd1, by Abcam (1 mentions) Ab74037, by Abcam (1 mentions) Axioskop 2 microscope, by Zeiss (1 mentions) Donkey anti rabbit igg594, by Thermo Fisher Scientific (1 mentions) Azure c500, by Azure Biosystems (1 mentions) Tnf α, by Abcam (1 mentions) Epac1, by Abcam (1 mentions) Tris glycine gel, by Thermo Fisher Scientific (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Occludin, by Abcam (1 mentions) Traf6, by Abcam (1 mentions) Irak m, by Abcam (1 mentions) Il 12p40, by Abcam (1 mentions)

5 protocols using tnfaip3

1

Histological Analysis of Gland Size

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The HE staining was mainly used to evaluate pathological progress. Three most representative ×100 fields per slide were selected and gland number in glandular epithelium was calculated. We observed the diameter of gland in Ctrl group is about 40–50 µm, then we calculated the percentage of gland (diameter > 50 µm) in total glands to evaluate the gland size. The IHC assay was performed as described before (14 (link)). Primary antibodies to PGR, CCND1, STMN1, PCNA and TNFAIP3 were purchased from Abcam. LGR5, VIM and KRT4 antibodies were purchased from Sigma-Aldrich. Phosphate buffered saline (PBS) was used as a negative control.
Cheng Y., Lv Q., Xie B., Yang B., Shan W., Ning C., Li B., Xie L., Gu C., Luo X., Chen X, & Zhu Q. (2017). Estrogen and high-fat diet induced alterations in C57BL/6 mice endometrial transcriptome profile. Endocrine Connections, 7(1), 36-46.
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2

Immunohistochemical Analysis of Psoriasis

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Formalin fixed, paraffin-embedded tissue slides obtained from psoriasis patients and normal controls were heated for 30 min at 60°C, rehydrated, and epitope retrieved with Tris-EDTA, pH 6. Slides were blocked, incubated with primary antibody APOBEC3 (LS-C98892-400; Lifespan bioscience), FOSL (A03927; Boster), IL-36G (sc-80056; Santa Cruz Biotechnology), TNFAIP3 (ab74037, Abcam), IL-17RC (LS-C400522, Lifespan bioscience), and IL-17RA (LS-C359381, Lifespan bioscience) overnight at 4 °C. Slides were then washed and incubated with Donkey anti-Rabbit IgG 594, Donkey anti-Mouse IgG 488, or Donkey anti-Rat IgG 594 (all from Invitrogen) for 1 h at room temperature. Slides were washed and prepared in mounting medium with 4',6-diamidino-2-phenylindole (DAPI) (VECTASHIELD Antifade Mounting Medium with DAPI, H-1200, VECTOR). Images were acquired using Zeiss Axioskop 2 microscope and analyzed by SPOT software 5.1. Images presented are representative of at least three experiments from separate donors.
Hughes T.K., Wadsworth MH I.I., Gierahn T.M., Do T., Weiss D., Andrade P.R., Ma F., de Andrade Silva B.J., Shao S., Tsoi L.C., Ordovas-Montanes J., Gudjonsson J.E., Modlin R.L., Love J.C, & Shalek A.K. (2020). Second-Strand Synthesis-Based Massively Parallel scRNA-Seq Reveals Cellular States and Molecular Features of Human Inflammatory Skin Pathologies. Immunity, 53(4), 878-894.e7.
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3

Western Blot Analysis of Cell Signaling

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Western blotting was performed as previously described [12 (link),13 (link)]. Briefly, cell lysates or whole retina lysates were separated onto precast tris-glycine gels (Invitrogen, Carlsbad, CA) and then blotted onto nitrocellulose membrane. After blocking in TBST, membranes were treated with Epac1, TNFAIP3, TRAF6, TNFα, phosphorylated and total NF-kB, phosphorylated and total IkB, occludin, ZO-1 (Abcam, Cambridge, MA), or beta actin (Santa Cruz Biotechnology, Santa Cruz, CA) primary antibodies. Secondary species-appropriate antibodies labeled with horseradish peroxidase were applied after the primary antibodies. Chemiluminescence (Thermo Scientific, Pittsburgh, PA) was used to visualize antigen–antibody complexes. Images were acquired on an Azure C500 (Azure Biosystems, Dublin, CA), and optical densities were measured using Image Studio Lite software (Lincoln, NE).
Liu L., Jiang Y, & Steinle J.J. (2022). TNFAIP3 is anti-inflammatory in the retinal vasculature. Molecular Vision, 28, 124-129.
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4

Spinal Cord and DRG Tissue Analysis

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This entailed the following: once the animals were anesthetized, 4% paraformaldehyde-based perfusion was done followed by sampling. The spinal cord and DRGs were embedded in paraffin and sliced as 10 micrometers thick sections employing a paraffin microtome. Subsequent toluidine blue staining entailed the use of the staining kit adhering to the prescribed protocols. Immunohistochemistry employed the use of TNFAIP3 (Rabbit, 1 : 200, Abcam, USA) followed by assessing and imaging in a confocal microscope.
Liu Y., Li J., Yao H., Zhang M., Lian J., Zhang H., Zhang K., Liu D., Chen J., Wang Y, & Gao Y. (2021). Effects of Upregulation of TNFAIP3 on Diabetic Neuropathic Pain in Mice. Disease Markers, 2021, 3470950.
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5

Macrophage Immune Response Profiling

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The levels of IL-12p40, IL-18, IL-23, TNFAIP3, IRAK-M, and Sp1 in whole-cell lysates of human macrophages were determined by ELISA (IL-12p40: Abcam, Cambridge, UK, IL-18: MBL life science, Woburn, MA, IL-23: Abcam; TNFAIP3: MYBioSource, San Diego, CA; IRAK-M, LifeSpan Biosciences, Seattle, WA; Sp1: Aviva Systems Biology Corporation, San Diego, CA) 6 h after exposure to LPS (10 ng/mL) or resiquimod (5μM) for 6 h. The levels of IL-23, IL-12p40, and IL-18 in whole-cell lysates prepared from human macrophages were determined by ELISA after sequential stimulation with LPS (0.1 ng/mL, 1 ng/mL, and 10 ng/mL for 6 h), Pam3CsK4 (1 μg/mL for 6 h) and then resiquimod (6 h). The sensitivity of ELISA for the various proteins was as follows: IL-12p40, 12.29 pg/mL; IL-18, 8.3 pg/mL; IL-23, 16.3 pg/mL; TNFAIP3, 23.5 pg/mL; IRAK-M, 0.78 ng/mL; and Sp1, 0.15 ng/mL.
Yamaguchi R., Sakamoto A., Yamaguchi R., Haraguchi M., Narahara S., Sugiuchi H, & Yamaguch Y. (2022). IL-23 production in human macrophages is regulated negatively by tumor necrosis factor α-induced protein 3 and positively by specificity protein 1 after stimulation of the toll-like receptor 7/8 signaling pathway. Heliyon, 8(2), e08887.
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