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spr experiment  (MedChemExpress)


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    MedChemExpress spr experiment
    CAM12203 directly binds <t>to</t> <t>DGK</t> ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by <t>SPR.</t> Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.
    Spr Experiment, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Macrophage DGK ζ -mediated phosphatidic acid remodeling aggravates acute liver failure"

    Article Title: Macrophage DGK ζ -mediated phosphatidic acid remodeling aggravates acute liver failure

    Journal: Acta Pharmaceutica Sinica. B

    doi: 10.1016/j.apsb.2025.06.019

    CAM12203 directly binds to DGK ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by SPR. Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.
    Figure Legend Snippet: CAM12203 directly binds to DGK ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by SPR. Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.

    Techniques Used: Concentration Assay, Binding Assay, Incubation, Western Blot



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    CAM12203 directly binds <t>to</t> <t>DGK</t> ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by <t>SPR.</t> Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.
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    CAM12203 directly binds <t>to</t> <t>DGK</t> ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by <t>SPR.</t> Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.
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    CAM12203 directly binds to DGK ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by SPR. Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.

    Journal: Acta Pharmaceutica Sinica. B

    Article Title: Macrophage DGK ζ -mediated phosphatidic acid remodeling aggravates acute liver failure

    doi: 10.1016/j.apsb.2025.06.019

    Figure Lengend Snippet: CAM12203 directly binds to DGK ζ . (A) Structure of the biotinylated probe. (B) RAW264.7 cells (pretreated with or without the probe for 2 h) were stimulated with LPS for 4 h. The mRNA level of Il1b was detected by Q-PCR. (C) BMDMs (pretreated with or without the probe for 2 h) were stimulated with LPS (4 h) + ATP (another 45 min). The IL-1 β concentration in culture medium (CM) was examined by HTRF. (D) Overall scheme of the experiments to identify the targets of CAM12203 (created with Biorender.com ). (E) The list of gene names of CAM12203 -binding proteins. (F) RAW264.7 lysate was pretreated with or without CAM12203 for 2 h (4 °C), followed by incubation with the probe for another 2 h (4 °C). The level of DGK ζ pulled down was determined by Western blotting. (G) RAW264.7 lysate was treated with or without CAM12203 , and the protein stability of DGK ζ resistant to pronase E was determined by DARTS. (H) RAW264.7 lysate was treated with or without CAM12203 , and the thermostability of DGK ζ was determined by CETSA. (I) DGK ζ protein was treated with CAM12203 , and the binding affinity was determined by SPR. Data are displayed as mean ± SEM ( n = 3). ∗ P < 0.05, ∗∗ P < 0.01 vs . the indicated group.

    Article Snippet: For the SPR experiment, 6 × His-tagged, recombinant full-length human DGK ζ (expressed in HEK293T cells, purified by Ni-NTA His-Tag purification agarose (HY–K0210, MedChemExpress)) was prepared.

    Techniques: Concentration Assay, Binding Assay, Incubation, Western Blot