mouse ccl2 elisa kit  (R&D Systems)

Journal: The FASEB Journal

Article Title: Lymphocyte Antigen 6G Mediates Vagotomy‐Associated Reduction in Body Weight

doi: 10.1096/fj.202600151RR

Figure Lengend Snippet: Vagotomy promoted Ly6G + cell infiltration into eWAT. Wild‐type mice were subjected to left cervical vagotomy (VX) or sham surgery and tissues were collected at 7 days following surgery. (A) The percentage of non‐adipocyte nuclei of total nuclei per section was quantified using ImageJ ( n = 4) and right panels show representative images of paraffin sections of eWAT stained with H&E in sham and VX animals. (B) CCL2 release from eWAT was analyzed by ELISA. The bar shows the CCL2 levels from sham ( n = 4) or VX ( n = 4) mice normalized to eWAT weight: ng/mL per g ± SEM (unpaired Student's t test). (C) eWAT was collected at 1 ( n = 3), 4 ( n = 4 sham, n = 5 VX), and 7 ( n = 15) days following VX or sham surgery and the eWAT SVCs were analyzed by flow cytometry. The bar shows the % ± SEM of CD11b + Ly6G + cells from CD45 + (one‐way ANOVA, Uncorrected Fisher's LSD). (D) Graphs show representative gating for CD11b + Ly6G + cells in sham and VX eWAT at 7 days (concatenated n = 5–6). (E) Representative immunostaining of Ly6G (red) and Perilipin1 (green) in paraffin sections of eWAT. (F–H) Bone marrow neutrophils after sham ( n = 9) or VX ( n = 5) surgery were isolated using negative magnetic beads and analyzed using bulk RNAseq (DESeq2). Heatmap (F), volcano plot (G) of differentially expressed genes, and GO (Gene Ontology) (H) enrichment bar plot. ns = not significant, * p < 0.05. VX, Vagotomy; eWAT, epididymal white adipose tissue; H&E, hematoxylin–eosin; SVCs, stromal vascular cells.

Article Snippet: In vitro release rate was calculated as mg NEFAs per mg eWAT tissue per hour. (2) CCL2 levels were quantified using a mouse CCL2 ELISA kit (R&D Systems, #DY497‐05), according to the manufacturer's instructions ( n = 1 experiment).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Immunostaining, Isolation, Magnetic Beads, RNA sequencing

Journal: The Journal of Clinical Investigation

Article Title: Characterization of intestinal immune responses in generalized human and murine lipodystrophy

doi: 10.1172/JCI192322

Figure Lengend Snippet: ( A ) Representative images of Pparg fl/fl Adipoq-Cre (Lipo) and WT mice showing the complete absence of fat tissue and H&E-stained images of mesenteric adipose tissue (arrows indicate mesenteric and gonadal fat tissue). Scale bars: 100 μm. ( B ) Leptin plasma levels (WT n = 4; Lipo n = 4). ( C ) Liver weights of WT mice ( n = 16) and Pparg fl/fl Adipoq-Cre mice ( n = 12) from 2 independent experiments with a representative image. ( D – G ) Flow cytometric assessment of splenic NK cells from Pparg fl/fl Adipoq-Cre mice ( n = 9–24) compared with WT littermate cells ( n = 10–23). Data were pooled from 3 independent experiments. ( H ) Representative H&E-stained images and immune cell counts (millions/cm) in the terminal ileum or colon as assessed by histology (WT: n = 12–16; Lipo: n = 8–12). Scale bars: 100 μm. ( I ) Gating strategy of T cells and CD4 + FoxP3 + Tregs. The line in the box indicates the median. Boxes range from the 25th to 75th percentiles. Whisker plots show the minimum (smallest) and maximum (largest) values while the line in the box indicates the median. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, by unpaired 2-tailed t test with Welch’s correction.

Article Snippet: Plasma levels of leptin or anti– E. coli (LPS) IgG were determined using the mouse Leptin DuoSet ELISA kit (R&D Systems) or the mouse anti– E. coli LPS (O111:B4) Antibody ELISA Kit (Chondrex).

Techniques: Staining, Clinical Proteomics, Whisker Assay

Journal: The Journal of Clinical Investigation

Article Title: Characterization of intestinal immune responses in generalized human and murine lipodystrophy

doi: 10.1172/JCI192322

Figure Lengend Snippet: ( A ) Experimental design: After 1 DSS cycle (1.5%), lipodystrophic or WT mice were transplanted with 500–600 mg adipose tissue from WT or leptin-deficient ob/ob donors by mini-laparotomy before undergoing another 2 cycles of DSS. Image on right shows vascularized transplanted fat 1 month after surgery. ( B ) Weight change of transplanted fat tissue relative to baseline. ( C ) Plasma leptin levels ( n = 4–9). ( D ) Representative images of H&E- stained colon sections from transplanted versus nontransplanted DSS-treated animals. Scale bars: 100 μm. ( E ) Box-and-whisker plots summarizing the histologic inflammation score of fat-transplanted and nontransplanted animals. Data shown were pooled from 2 independent transplantation experiments (bold symbols) and additional control data points derived from nontransplantation DSS experiments (light gray) shown in ( n = 4–18). ( F ) Liver weights of transplanted WT and Pparg fl/fl Adipoq-Cre mice ( n = 4–18; data were pooled from 5 experiments). ( G ) Representative FACS plots showing IFN-γ and IL-17A production in colonic CD4 + T cells. UNSTIM., unstimulated. ( H ) Box-and-whisker plots summarizing absolute numbers of IFN-γ– and IL-17A–producing CD4 + T cells normalized to WT mice ( n = 4–18; data were pooled from 5 experiments). Statistical differences were calculated by 1-way ANOVA with Šídák’s correction. Each point represents 1 mouse; boxes range from the 25th-75th percentiles. Whisker plots show the minimum (smallest) and maximum (largest) values while the line in the box indicates the median. ( I ) Experimental setup and representative FACS plots showing IFN-γ– and IL-17A–producing CD4 + T cells in peripheral blood of a patient with AGLCD before and 4 days after daily recombinant leptin substitution. transpl., transplantation. Data indicate the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, by 1-way ANOVA with Tukey’s multiple comparisons test for B , C , E , F , and H .

Article Snippet: Plasma levels of leptin or anti– E. coli (LPS) IgG were determined using the mouse Leptin DuoSet ELISA kit (R&D Systems) or the mouse anti– E. coli LPS (O111:B4) Antibody ELISA Kit (Chondrex).

Techniques: Clinical Proteomics, Staining, Whisker Assay, Transplantation Assay, Control, Derivative Assay, Recombinant

Journal: Molecular Nutrition & Food Research

Article Title: Bovine Milk Extracellular Vesicles Modulate Alveolar Bone Microarchitecture and Mitigate Hepatic Steatosis in Obese Mice Fed a High‐Fat Diet

doi: 10.1002/mnfr.70400

Figure Lengend Snippet: Analysis of systemic bone remodeling markers of mice by ELISA. (A) RANKL, (B) OPG, and (C) RANKL/OPG ratio in serum of mice fed a control (C) diet, or a high‐fat (HF) diet for 12 weeks and treated with bovine MEVs in the last 4 weeks ( n = 5–6 per group). Bars represent mean values ± standard error of the mean. Statistical difference represented by * p < 0.05 ‐ HF versus C and # p < 0.05 ‐ HF versus MEVs, one‐way ANOVA, Dunnett posttest for all data.

Article Snippet: The bone metabolism markers RANKL and OPG, as well as metabolic adipose serum markers such as adiponectin and leptin, were quantified in mouse serum using DuoSet ELISA kits (R&D Systems Europe Ltd., Abington, United Kingdom).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular Nutrition & Food Research

Article Title: Bovine Milk Extracellular Vesicles Modulate Alveolar Bone Microarchitecture and Mitigate Hepatic Steatosis in Obese Mice Fed a High‐Fat Diet

doi: 10.1002/mnfr.70400

Figure Lengend Snippet: Analysis of weight, body composition, and metabolic alterations of mice. (A) Final weight, (B) serum Leptin by ELISA, (C) weight of epididymal adipose tissue (EAT), (D) EAT adipocyte area by histomorphometry analysis, (E) stratification of the EAT adipocyte area, (F) representative image of EAT adipocyte area (x100), (G) weight of subcutaneous adipose tissue (SAT), (H) SAT adipocyte area by histomorphometry analysis, (I) representative image of SAT adipocyte area (x100), and (J) stratification of the SAT adipocyte area. Histological scale bars, 100 µm. (K) serum Adiponectin by ELISA, (L) serum glucose, (M) oral glucose tolerance test (OGTT), (N) area under the curve (AUC)—OGTT, (O) serum triglycerides, and (P) serum total cholesterol of mice fed a control (C) diet or a high‐fat (HF) diet for 12 weeks and treated with bovine MEVs in the last 4 weeks ( n = 6 per group). Bars represent mean values ± standard error of the mean. Statistical difference represented by * p < 0.05 ‐ HF versus C and # p < 0.05 ‐ HF versus MEVs, one‐way ANOVA, Dunnett posttest for all data.

Article Snippet: The bone metabolism markers RANKL and OPG, as well as metabolic adipose serum markers such as adiponectin and leptin, were quantified in mouse serum using DuoSet ELISA kits (R&D Systems Europe Ltd., Abington, United Kingdom).

Techniques: Enzyme-linked Immunosorbent Assay, Control