Tapi 0

Manufactured by Santa Cruz Biotechnology

Sourced in United States

TAPI-0 is a protein inhibitor that selectively targets and inhibits the activity of the TACE (Tumor Necrosis Factor-Alpha Converting Enzyme) enzyme. TACE plays a crucial role in the proteolytic cleavage and release of various membrane-bound proteins, including the pro-inflammatory cytokine TNF-alpha. By inhibiting TACE, TAPI-0 can modulate the levels of these important signaling molecules.

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Lab products found in correlation

Tnf α, by Enzo Life Sciences (1 mentions) Anti tlr4 antibody, by Santa Cruz Biotechnology (1 mentions) Bq 123, by MedChemExpress (1 mentions) Tak 242, by Merck Group (1 mentions) Anti cd8, by BD (1 mentions) Anti cd69, by BioLegend (1 mentions) Live dead fixable near infra red dead cell stain, by Thermo Fisher Scientific (1 mentions) Anti tnf, by BD (1 mentions) Facsaria fusion, by BD (1 mentions) Anti cd3, by BD (1 mentions) Anti cd4, by BD (1 mentions)

3 protocols using tapi 0

Investigating ADAM17 Variants in APP-Expressing SH-SY5Y Cells

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SH-SY5Y cells stably expressing human APP₆₉₅ carrying the Swedish double mutation (in pcDNA3.1, selection with 0.3 mg/ml hygromycin) were kindly provided by Tobias Hartmann and Marcus Grimm (Saarland University). SH-SY5Y cells were stably transfected with expression constructs for ADAM17 variants (in pcDNA3.1, selection with 500 μg/ml geneticin). Cells were grown in DMEM with selection antibiotics and 10% fetal calf serum (FCS). For rescue experiments, cells were supplied with media containing 100 nM TAPI-0 (SantaCruz) or equal volume of DMSO as control. Medium was changed every second day, cells were checked for mycoplasma contamination.

Hartl D., May P., Gu W., Mayhaus M., Pichler S., Spaniol C., Glaab E., Bobbili D.R., Antony P., Koegelsberger S., Kurz A., Grimmer T., Morgan K., Vardarajan B.N., Reitz C., Hardy J., Bras J., Guerreiro R., Balling R., Schneider J.G, & Riemenschneider M. (2018). A rare loss-of-function variant of ADAM17 is associated with late-onset familial Alzheimer disease. Molecular Psychiatry, 25(3), 629-639.

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Reagents for In Vitro Inflammatory Studies

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BIM, bovine serum albumin (BSA), BQ-788, 4′,6-diamidino-2-phenylindole (DAPI), ET-1, Ficoll^® PM 400 (Ficoll), LPS from Escherichia coli 0111:B4, TAK-242, and WP9QY were purchased from Merck (Darmstadt, Germany). H398 and TNF-α were obtained from Enzo Life Science (Farmingdale, NY, USA) and FUJIFILM Wako Chemicals (Tokyo, Japan), respectively. TAPI-0 and anti-TLR4 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). BQ-123 and PA were obtained from MedChemExpress (Monmouth Junction, NJ, USA) and AG Scientific (San Diego, CA, USA), respectively. L-NMMA was purchased from Dojindo (Kumamoto, Japan). NBD-CSA was synthesized following our previous report by Tajima et al. [7 (link)]. All other reagents were commercially obtained.

Daikohara K., Akanuma S.I., Kubo Y, & Hosoya K.I. (2022). Lipopolysaccharide-Induced Functional Alteration of P-glycoprotein in the Ex Vivo Rat Inner Blood–Retinal Barrier. International Journal of Molecular Sciences, 23(24), 15504.

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Characterization of Antigen-Specific CD8+ T Cells

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Antigen-specific CD8⁺ T cells were identified and viable cells sorted using surface-trapped TNF-α staining (23 (link)). PBMC, isolated CD8β+ T cells, or CD8⁺ T cell transductants were incubated with peptide-pulsed antigen-presenting cells, the co-stimulatory molecules CD28 and CD49d (BD Biosciences), anti-TNF (clone: MAb11, PE, BD Biosciences) and TAPI-0 (5uM final concentration, Santa Cruz Biotechnology). Cells were incubated at 37°C for 8 hours. After incubation, cells were stained anti-CD3 (clone: SP34–2, Pacific Blue, BD Biosciences), anti-CD8 (clone: SK1, TruRed, BD Biosciences), anti-CD4 (clone: L200, PE-Cy7, BD Biosciences), anti-CD14 (clone: M5E2, APC, Biolegend), anti-CD69 (FN50, FITC, BioLegend), anti-CD16 (clone: 3G8, APC, Biolegend), and anti-CD20 (clone: 2H7, APC, Biolegend) and LIVE/DEAD Fixable Near Infra-Red Dead Cell Stain (Life Technologies) was used to assess cell viability. Antigen-specific cells were defined as TNF+/CD69+ CD8⁺ T cell responses 2x the magnitude of the no peptide control, with the no peptide control responses below 0.5%. Viable antigen-specific cells were sorted using a FACSAria Fusion (BD Biosciences), and analysis was conducted with FlowJo software (Tree Star).

Abdulhaqq S., Ventura A.B., Reed J.S., Bashirova A.A., Bateman K.B., McDonald E., Wu H.L., Greene J.M., Schell J.B., Morrow D., Wisskirchen K., Martin J.N., Deeks S.G., Carrington M., Protzer U., Früh K., Hansen S.G., Picker L.J., Sacha J.B, & Bimber B.N. (2021). Identification and characterization of antigen-specific CD8+ T cells using surface-trapped TNF-α and single-cell sequencing. Journal of immunology (Baltimore, Md. : 1950), 207(12), 2913-2921.

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