human ccl5 elisa kit  (Elabscience Biotechnology)

Journal: iScience

Article Title: Serglycin modulates inflammation and metabolism in macrophages

doi: 10.1016/j.isci.2026.115235

Figure Lengend Snippet: SRGN deficiency reshapes the inflammatory secretome and transcriptional programs in human macrophages (A) Volcano plot of differentially secreted proteins between SRGN −/− and wild-type THP-1 macrophages under M1 polarization. Among 1,507 quantified proteins, 53 were significantly altered (adjusted p < 0.05), with serglycin being the most downregulated protein in knockout cells. (B) Validation of selected targets by RT-qPCR. SRGN −/− M1 macrophages showed significantly increased expression of IL6 and TNF and reduced expression of CCL5 compared with wild-type cells (mean ± SEM; unpaired Student’s t test; p < 0.05, ∗ p < 0.01, and ∗∗∗ p < 0.0001; n = 4). (C) ELISA quantification of secreted cytokines in culture supernatants. SRGN −/− macrophages secreted significantly less TNF-α, CCL5, and IL-6 compared with wild-type macrophages ( n = 4), consistent with proteomics and RNA-seq data. (D) Transmission electron microscopy (TEM) images of THP-1 M0 and M1 macrophages. Scale bars, 5 μm. Vesicles were manually annotated and quantified in 10 cells per experimental group. The number of vesicles per cell and the percentage of cellular area occupied by vesicles were significantly reduced in both M0 and M1 SRGN −/− macrophages compared with wild-type macrophages. (E) Phagocytosis assay using fluorescently labeled bioparticles. SRGN −/− macrophages exhibited reduced phagocytic capacity under both M0 and M1 conditions (mean ± SEM; unpaired Student’s t test; p < 0.05 and ∗∗ p < 0.001; n = 6).

Article Snippet: THP-1: cytokine levels were measured using ELISA kits from R&D Systems: CCL5 (Cat. No. DY278), IL6 (Cat. No. DY206), IL-1β (Cat. No. DY201), TGF-β (Cat. No. DY240; activation kit DY010), and TNF-α (Cat. No. DY210), following the manufacturer’s instructions.

Techniques: Knock-Out, Biomarker Discovery, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Transmission Assay, Electron Microscopy, Phagocytosis Assay, Labeling

Journal: Journal of Advanced Research

Article Title: CCR1 hi /CCL5 hi macrophage-mediated CCL5 hi T cell chemotaxis in salivary gland aggravates Sjögren’s syndrome

doi: 10.1016/j.jare.2025.06.076

Figure Lengend Snippet: CCR1-overexpressing macrophages induce T and B cell migration. (A) Immunofluorescence (IF) detection of CD68 expression in THP-1 cells induced by PMA (100 nM, 24 h). (B) Quantitative analysis of CD68 expression based on IF staining fluorescence intensity. (C) Schematic diagram of the detection of macrophage-induced T and B cell migration. (D, E) Western blot analysis and quantitative evaluation of CCR1 and CCL5 expression in macrophages after transfection with the CCR1 overexpression plasmid. (F) ELISA measurement of CCL5 concentration in supernatants from CCR1-overexpressing macrophages. (G, H) Transwell assay evaluating macrophage-mediated migration of T and B cells, with quantitative analysis of migrated cell numbers. Statistical significance was determined using one-way ANOVA.

Article Snippet: The release level of CCL5 in cell culture supernatants was assessed using a commercial ELISA kit (EK1129, Lianke Biotechnology Co., Ltd., Hangzhou, China) following the manufacturer’s instructions.

Techniques: Migration, Immunofluorescence, Expressing, Staining, Fluorescence, Western Blot, Transfection, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay

Journal: Frontiers in Immunology

Article Title: From multi-omics to functional validation: the PTMRS stratifies TME and positions PDGFRB in CRC biology

doi: 10.3389/fimmu.2025.1728291

Figure Lengend Snippet: Impact of PDGFRb perturbation within tumor-associated fibroblasts on CRC cell phenotypes. (A) Western blots of t-PDGFRb and p-PDGFRb in CCD-18Co fibroblasts treated with vehicle (Veh), PDGF-BB, or PDGF-BB plus SUN. b-actin serves as a loading control. (B) Quantification of the p-PDGFRb/t-PDGFRb ratio relative to the Veh group. (C) Representative EdU staining of HCT116 cells cultured with CM from the three fibroblast conditions (Veh-CM, PDGF-BB-CM, SUN-CM). EdU (green), DAPI (blue), merged images as indicated. (D) Relative EdU labeling index in HCT116 cells. (E, F) CCK-8 assays showing relative cell viability of HCT116 (E) and DLD-1 (F) cells after 48 h exposure to Veh-CM, PDGF-BB-CM, or SUN-CM. (G) Representative Transwell images of migrated HCT116 and DLD-1 cells under the indicated CM conditions. (H, I) Quantification of migrated HCT116 (H) and DLD-1 (I) cells per 10× field. (J) Violin plots showing concentrations of CCL5, CXCL12, IL-6, IL-10, and TGF-b1 in fibroblast CM measured by ELISA. *P < 0.05, **P < 0.01, ***P < 0.001 vs Veh group; ns, no significant difference. Abbreviations: Veh, vehicle; SUN, sunitinib; CM, conditioned medium; PDGF-BB, the BB homodimer of platelet-derived growth factor.

Article Snippet: Concentrations of C-C motif chemokine ligand 5 (CCL5), C-X-C motif chemokine ligand 12 (CXCL12), interleukin-6 (IL-6), interleukin-10 (IL-10), and transforming growth factor-β1 (TGF-β1) were measured in the conditioned media(CM) of human colonic fibroblasts CCD-18Co (processed, collected as described above, and stored at −80 °C) using human enzyme-linked immunosorbent assay (ELISA) kits from R&D Systems (CCL5, Catalog #DRN00B; CXCL12, Catalog #DSA00; IL-6, Catalog #D6050B; IL-10, Catalog #D1000B; TGF-β1, Catalog #DB100C).Recombinant cytokine standards provided with each kit were serially diluted to generate standard curves.

Techniques: Western Blot, Control, Staining, Cell Culture, Labeling, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay