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  • notch1  (ATCC)
    93
    ATCC notch1
    (A) BMMs were stimulated by LPS/IFNγ for indicated times in the presence of DMSO vehicle control or GSI (25 μM) as described in the materials and methods. Total cell lysates were harvested and analyzed for <t>Notch1</t> and cleaved Notch1 (Val1744) using Western blotting. The results represent three independent experiments. (B-C) BMMs were activated as described in (A) without GSI for indicated times, and the expression levels of Hes1 and Hes5 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (D-E) BMMs were activated as described in (A) for 4 hrs, and the expression levels of il12p40 and il23p19 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (*) indicates where p < 0.05, which was considered to be statistically significant. (F) BMMs were pretreated with GSI (25 μM) or vehicle control DMSO and activated for 24 hr to become CA and regulatory macrophages as described above. The amount of IL-12p70 was measured in the culture supernatants using ELISA. The results are the mean±SD and represent two independent experiments. ND = not detectable.
    Notch1, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) BMMs were stimulated by LPS/IFNγ for indicated times in the presence of DMSO vehicle control or GSI (25 μM) as described in the materials and methods. Total cell lysates were harvested and analyzed for Notch1 and cleaved Notch1 (Val1744) using Western blotting. The results represent three independent experiments. (B-C) BMMs were activated as described in (A) without GSI for indicated times, and the expression levels of Hes1 and Hes5 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (D-E) BMMs were activated as described in (A) for 4 hrs, and the expression levels of il12p40 and il23p19 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (*) indicates where p < 0.05, which was considered to be statistically significant. (F) BMMs were pretreated with GSI (25 μM) or vehicle control DMSO and activated for 24 hr to become CA and regulatory macrophages as described above. The amount of IL-12p70 was measured in the culture supernatants using ELISA. The results are the mean±SD and represent two independent experiments. ND = not detectable.

    Journal: Molecular Immunology

    Article Title: Involvement of Notch Signaling Pathway in Regulating IL-12 Expression via c-Rel in Activated Macrophages

    doi: 10.1016/j.molimm.2012.03.017

    Figure Lengend Snippet: (A) BMMs were stimulated by LPS/IFNγ for indicated times in the presence of DMSO vehicle control or GSI (25 μM) as described in the materials and methods. Total cell lysates were harvested and analyzed for Notch1 and cleaved Notch1 (Val1744) using Western blotting. The results represent three independent experiments. (B-C) BMMs were activated as described in (A) without GSI for indicated times, and the expression levels of Hes1 and Hes5 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (D-E) BMMs were activated as described in (A) for 4 hrs, and the expression levels of il12p40 and il23p19 were measured using qPCR. The results are the mean±SD and represent two independent experiments. (*) indicates where p < 0.05, which was considered to be statistically significant. (F) BMMs were pretreated with GSI (25 μM) or vehicle control DMSO and activated for 24 hr to become CA and regulatory macrophages as described above. The amount of IL-12p70 was measured in the culture supernatants using ELISA. The results are the mean±SD and represent two independent experiments. ND = not detectable.

    Article Snippet: 2.6 Overexpression of Truncated Intracellular Notch1 Overexpression of the truncated intracellular form of Notch1 (N IC ) in RAW 264.7 cells (ATCC No. TIB-71) was carried out using pcDNA3 plasmid containing the intracellular Notch1 (N IC )-encoding sequences corresponding to amino acid residues 1759–2556 (pcDNA3N IC ), and empty pcDNA3 plasmid was used as a control vector (both were kind gifts from Professor Barbara Osborne, University of Massachusetts at Amherst, USA).

    Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay