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Biacore spr experiments
Spr Experiments, supplied by Biacore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Scaffold-hopping for molecular glues targeting the 14-3-3/ERα complex
Article Snippet: The SPR experiments were performed at 25 °C using a Biacore X100 (Cytiva) and a 200 nm Strep-Tactin XT derivatized linear polycarboxylate hydrogel chip, medium charge density (XanTec Bioanalytics).

Article Title: Scaffold-hopping for molecular glues targeting the 14-3-3/ERα complex.
Article Snippet: The assay was performed in 384- well microplates (Corning 4513, low volume white) at a volume of 10μL per well.

Article Title: A Surface Plasmon Resonance-Based Integrated Assay for Quantification and Glycosylation Characterization of Monoclonal Antibodies in Crude Heterogeneous Samples.
Article Snippet: All SPR experiments described in this manuscript were performed on a Biacore T100 biosensor using a CM5 sensor chip (Cytiva) and HBS‐EP as running buffer.

Article Title: Discovery of novel non-sulfonylurea NLRP3 inflammasome inhibitors for the treatment of multiple inflammatory diseases.
Article Snippet: Targeting NLRP3 inflammasome has emerged as a promising therapeutic strategy for various inflammatory diseases.. In this work, to discover safe and effective novel NLRP3 inhibitors, we designed and synthesized a series of new non-sulfonylurea NLRP3 inflammasome inhibitors.. Among them, the representative compound B6 specifically and potently inhibited the activation of NLRP3 inflammasome with an IC50 of 10.69 nM, exhibiting better potency compared to MCC950 (IC50 = 14.08 nM).

Article Title: Designed miniproteins potently inhibit and protect against MERS-CoV
Article Snippet: SPR experiments were performed using a Biacore 8K and the biotin CAP chip (Cytiva) for both the monomeric and trimeric designs.

Article Title: Understanding the mode of action of BtEcR/USP with benzpyrimoxan in combination with high throughput SPR screening and molecular simulation approaches
Article Snippet: Bemisia tabaci (B. tabaci) is a major agricultural pest that infests over 500 plant species, posing a significant threat to agricultural production due to its polyphagy, adaptability, and ability to transmit plant viruses.. Excessive and improper insecticide use has caused resistance to pyrethroids, organophosphates, and neonicotinoids, creating an urgent need for new insecticides with novel structures and mechanisms of action.. In this study, we developed an in vitro test platform targeting B. tabaci ecdysteroid receptor (BtEcR/USP-LBD) using Surface Plasmon Resonance (SPR) and investigated the novel insect growth regulator benzpyrimoxan through SPR, molecular docking, and molecular dynamics (MD) simulations.

Article Title: Architecture and conformational dynamics of the BAM-SurA holo insertase complex.
Article Snippet: SPR experiments were conducted on a Biacore T100 (GE Healthcare).

Article Title: Paeonol ameliorates diabetic nephropathy by promoting TFEB-mediated lysosome biogenesis and lipophagy.
Article Snippet: Rationale: Diabetic nephropathy (DN) is a significant clinical and public health burden worldwide whose magnitude underscores the urgent need for more effective treatment options.. Excessive lipid accumulation in renal tubular cells leads to their injury and dysfunction, thereby contributing to DN progression, suggesting that the alleviation of renal tubular lipid accumulation is a potential strategy for treating DN.. Methods: The effects of paeonol (PAE), a natural phenolic compound, on renal tubular lipid accumulation were evaluated using a glucolipotoxicity-treated HK-2 cell line and C57BL/6 J mice treated with streptozotocin (STZ) injection combined with a high-fat diet (HFD).



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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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Biacore spr binding experiments
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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Cytiva Europe surface plasmon resonance (spr) spr binding experiments
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.
Surface Plasmon Resonance (Spr) Spr Binding Experiments, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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