duoset elisa development systems canine kits  (R&D Systems)

Journal: Parasite Immunology

Article Title: The Regulatory Effect of Histone Deacetylase ( HDAC ) 1 and 2 on iNOS , IL ‐6, TNF ‐α and IL ‐10 Expression in Canine Macrophages Infected With Leishmania infantum

doi: 10.1111/pim.70078

Figure Lengend Snippet: Inhibition of HDAC1 in cells infected with Leishmania infantum and treated with sodium butyrate on IL‐6, TNF‐α and IL‐10 expression. DH82 cells were treated with the pharmacological HDAC1 inhibitor sodium butyrate NaB (10 mM) and (20 mM). After 6 h of infection, the culture supernatant cells were harvested, and (A) IL‐6, (B) TNF‐α and (C) IL‐10 were measured by capture ELISA. The statistical test used was the Friedman test, followed by Dunn's multiple‐comparison test ( p < 0.05).

Article Snippet: Cytokines IL‐6, TNF‐α and IL‐10 were quantified in culture supernatant from L. infantum infected macrophages after HDAC1 and HDAC2 suppression with NaB (10 mM) and (20 mM) (Sigma‐Aldrich, MO, USA) by using the DuoSet ELISA Development Systems canine kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer's instructions.

Techniques: Inhibition, Infection, Expressing, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Breast Cancer Research : BCR

Article Title: IL-3/STAT5/miR-155-5p axis supports stem-related pathway reprogramming in TNBC

doi: 10.1186/s13058-025-02143-1

Figure Lengend Snippet: IL-3, STAT5 and miR-155-5p expression based on TCGA or GEO database and miR-155-5p expression upon IL-3-mediated STAT5A activation. ( A ) IL-3 expression in TNBC compared to non-TNBC samples based on GEO dataset. ( B-C ) IL-3 expression in TNBC compared to non-TNBC and normal breast samples on TCGA dataset ( D ) ELISA assay to evaluate the release of IL-3 in the supernatant of 7 different breast cancer cell lines (n = 3; 3 biological replicates for each n). ( E , F ) RNAseq analysis of miR-155-5p expression in TNBC compared to non-TNBC samples and normal tissue based on TCGA dataset. ( G ) Spearman correlation analysis between the expression of IL-3 and miR-155-5p. ( H ) RT-PCR analysis of miR-155-5p expression in MDA-MB-436 and Hs-578T cells treated with IL-3 (5 ng/ml) for 6h and 16h. RNU6B was used as internal control (n = 6; 6 biological replicates for each n). ( I , J ) Spearman correlation analysis between the expressions of STAT5A and miR-155-5p and miR-155-3p. ( K ) Western Blot analysis of p-STAT5 (Tyr694) in MDA-MB-436 and Hs-578T cells treated with IL-3 (5 ng/ml) for 0, 10, 20 and 30 min. M07 cells treated with IL-3 were used as p-STAT5A internal control. Total form of STAT5 was used as phosphorylation control and β-actin was used as housekeeping loading control (n = 3; 3 biological replicates for each n). ( L ) RT-PCR results from chromatin immunoprecipitation of the BIC promoter via pSTAT5A binding following IL-3 treatment (5 ng/ml) for 10 min in MDA-MB-436 and Hs-578T TNBC cell lines. (n = 3; 3 biological replicates for each n). The data are represented as mean ± SEM

Article Snippet: Relative quantification of human IL-3 secretion was performed using DuoSet ELISA Development Systems (R&D Systems #DY007B) according to the manufacturer’s instructions.

Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Phospho-proteomics, Chromatin Immunoprecipitation, Binding Assay

Journal: Breast Cancer Research : BCR

Article Title: IL-3/STAT5/miR-155-5p axis supports stem-related pathway reprogramming in TNBC

doi: 10.1186/s13058-025-02143-1

Figure Lengend Snippet: IL-3 release is increased in 3D culture and partially relies on miR-155-5p ( A , E ) ELISA assay was performed to evaluate IL-3 release by MDA-MB-436 and Hs-578 T cells cultured in adherent (2D) and non-adherent (3D) condition (mammospheres) (n = 3; 3 biological replicates for each n). ( B , F ) ELISA assay to evaluate IL-3 release upon treatment for 24 h with saline, antagomiR-155 (10 nmol), NC (10 nmol), and IL-3 blocking mAb (1 ng/ml) (n = 3; 3 biological replicates for each n). ( C , G ) H&E staining from MDA-MB-436 and Hs-578 T–derived xenografts. ( D , H ) Representative images and quantification of IL-3 + area. Original magnification corresponds to 20X, scale bar to 50 µm. (n = 3; 3 biological replicates for each n). Data are represented as mean ± SEM

Article Snippet: Relative quantification of human IL-3 secretion was performed using DuoSet ELISA Development Systems (R&D Systems #DY007B) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Saline, Blocking Assay, Staining, Derivative Assay