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    GeneTex anti ho 1
    Generation of intracellular ROS promotes MCL-induced apoptosis ( A ) ROS levels in primary AML cells treated with 10 μM MCL at various time points over 240 min as measured by DCF-DA fluorescence. ( B ) ROS levels in primary CD34 + LSPCs treated with MCL for 1 h. ( C ) ROS levels in primary AML cells co-treated with MCL and NAC. ( D ) Percentage of viable LSPCs pretreated with NAC and then exposed to MCL. ( E ) Relative <t>HO-1</t> expression in primary AML cells treated with 10 μM MCL after 2 h and 6 h. Control represents untreated cells. Expression was normalized to GAPDH. ( F ) Western blot analysis of HO-1 protein expression in primary AML cells treated with 10 μM MCL for 6 h. GAPDH was used as a loading control. ( G ) Immunofluorescence assays that show cellular localization of HO-1 and Nrf2 in bone marrow cells isolated from AML mice. HO-1/Nrf2 proteins are shown in green and nuclei are shown in red. ( H ) Western blot analysis of p65 protein expression with or without ROS inhibitor NAC. ( I ) Mechanism of MCL in inducing cell apoptosis was summarized.
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    Generation of intracellular ROS promotes MCL-induced apoptosis ( A ) ROS levels in primary AML cells treated with 10 μM MCL at various time points over 240 min as measured by DCF-DA fluorescence. ( B ) ROS levels in primary CD34 + LSPCs treated with MCL for 1 h. ( C ) ROS levels in primary AML cells co-treated with MCL and NAC. ( D ) Percentage of viable LSPCs pretreated with NAC and then exposed to MCL. ( E ) Relative HO-1 expression in primary AML cells treated with 10 μM MCL after 2 h and 6 h. Control represents untreated cells. Expression was normalized to GAPDH. ( F ) Western blot analysis of HO-1 protein expression in primary AML cells treated with 10 μM MCL for 6 h. GAPDH was used as a loading control. ( G ) Immunofluorescence assays that show cellular localization of HO-1 and Nrf2 in bone marrow cells isolated from AML mice. HO-1/Nrf2 proteins are shown in green and nuclei are shown in red. ( H ) Western blot analysis of p65 protein expression with or without ROS inhibitor NAC. ( I ) Mechanism of MCL in inducing cell apoptosis was summarized.

    Journal: Oncotarget

    Article Title: Antineoplastic effects and mechanisms of micheliolide in acute myelogenous leukemia stem cells

    doi: 10.18632/oncotarget.11342

    Figure Lengend Snippet: Generation of intracellular ROS promotes MCL-induced apoptosis ( A ) ROS levels in primary AML cells treated with 10 μM MCL at various time points over 240 min as measured by DCF-DA fluorescence. ( B ) ROS levels in primary CD34 + LSPCs treated with MCL for 1 h. ( C ) ROS levels in primary AML cells co-treated with MCL and NAC. ( D ) Percentage of viable LSPCs pretreated with NAC and then exposed to MCL. ( E ) Relative HO-1 expression in primary AML cells treated with 10 μM MCL after 2 h and 6 h. Control represents untreated cells. Expression was normalized to GAPDH. ( F ) Western blot analysis of HO-1 protein expression in primary AML cells treated with 10 μM MCL for 6 h. GAPDH was used as a loading control. ( G ) Immunofluorescence assays that show cellular localization of HO-1 and Nrf2 in bone marrow cells isolated from AML mice. HO-1/Nrf2 proteins are shown in green and nuclei are shown in red. ( H ) Western blot analysis of p65 protein expression with or without ROS inhibitor NAC. ( I ) Mechanism of MCL in inducing cell apoptosis was summarized.

    Article Snippet: Cells were incubated in blocking buffer [10% FBS/0.1% Tween 20 in 1× PBS (pH 7.4)] and stained with rabbit polyclonal anti-p65 (C-20), anti-Nrf-2 (C-20) (Santa Cruz Biotechnology, Santa Cruz, CA), or anti-HO-1 (H-105; GeneTex Inc., San Antonio, TX) antibodies in 0.5% Triton X-100 for 2 h at RT.

    Techniques: Fluorescence, Expressing, Western Blot, Immunofluorescence, Isolation, Mouse Assay