mouse specific sandwich elisa kit  (Elabscience Biotechnology)

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: Transcriptome results showed that genes such as LCN2 and C1q were significantly upregulated in the hippocampus. A Volcano plot of differentially expressed genes (DEGs) in hippocampus from silica-exposed (Sil) versus vehicle (Veh) mice. Cut-off: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\:\left|{log}_{2}FC\right|\geqq\:O.O5$$\end{document} , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\:Pvalue\leqq\:0.05$$\end{document} , indicated by gray dashed lines in the plot. Red, significantly up-regulated; blue, significantly down-regulated; grey, non-significant. n = 3 biological replicates per group. B Gene set enrichment analysis of the 2,275 up-regulated genes using the Metascape database. C Inflammatory module network analysis of the DEGs enriched in GO:0006954 (GO database), circled by red in ( B ). D Neutrophil degranulation cascade module network analysis of the DEGs enriched in R-MMU-6,798,695 (Reactome database), circled by green in ( B ). E Microglial phagocytosis module network analysis of the DEGs enriched in WP3626 (WikiPathways database), circled by yellow in ( B ). F LCN2 and complement receptor C3ar1 were identified as having potential interactions. Interactions between genes among C-F are indicated by lines, whose colors represent interaction types from STRING database

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: LCN2 further activates microglia, exacerbating neuroinflammation in the hippocampus. A Representative immunohistochemical staining of LCN2 in the CA1, CA3, and DG regions of paraffin-embedded tissue from Veh and Sil groups; scale bar: 50 μm. The black arrow indicates LCN2-positive cells, brown DAB precipitate indicates positive immunoreactivity. B Quantitative analysis of positive staining from panel A, ( n = 4). C Western blot analysis assessing LCN2 protein expression. D LCN2 expression levels in Veh and Sil group mice ( n = 9). E Representative images of LCN2 immunoreactivity (green) and Iba-1 (red) in the CA1, CA3, and DG regions of frozen tissue from Veh and Sil groups; scale bar: 50 μm. F Representative immunofluorescence images of frozen sections showing Iba-1-positive cells (red); scale bar: 50 μm. G Sholl analysis of microglia in frozen tissue from Veh and Sil groups, measuring the distance from the soma (in 2 μm increments) and the number of microglial processes intersecting concentric circles ( n = 4). Maxima for all measures were observed approximately 20 μm from the soma. Data are presented as mean ± SEM. For panels ( B ) and ( D ), data were analyzed by t-tests. For panel ( G ), data were analyzed by two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns = not significant

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques: Immunohistochemical staining, Staining, Western Blot, Expressing, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: LCN2 activates microglia through the MC4R receptor rather than the MC1R or MC3R receptors in the hippocampus. A Western blot analysis assessing MC4R, MC1R, and MC3R protein expression. B Protein expression (normalized to β-actin) of MC4R, MC1R, and MC3R in tissue from Veh and Sil group mice ( n = 9). C Representative images of MC4R immunoreactivity (green), LCN2 immunoreactivity (red), and Iba-1 (purple) in frozen tissue. Scale bar: 50 μm. D Quantitative analysis of the MC4R-positive cells of CA1, CA3, and DG regions ( n = 4). E Quantitative analysis of the LCN2-positive cells of CA1, CA3, and DG regions ( n = 4). F Quantitative analysis of the MC4R + LCN2 + Iba-1 + cells (cell/mm 2 ) in the CA1, CA3, and DG regions ( n = 4). Data are presented as mean ± SEM. For panels (B-C, E-F), data were analyzed by t-tests. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns = not significant

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques: Western Blot, Expressing

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: LCN2-mediated cAMP/PKA/NF-κB signaling and C1q upregulation, with HS024 inhibiting C1q release in the BV2 cell culture. A ELISA measurement of cAMP in cell culture supernatant ( n = 4). The main effects of LCN2 (F = 92.206, p < 0.001) and HS024 (F = 18.470, p = 0.001) were both significant. B Western blot analysis assessing the expression levels of MC4R, PKA, NF-κB p65, and C1q in four groups. C Quantitative analysis of MC4R protein expression (normalized to β-actin) ( n = 3). The main effects of LCN2 (F = 6.863, p = 0.031) and HS024 (F = 17.455, p = 0.003) were significant. D Quantitative analysis of PKA protein expression (normalized to GAPDH) ( n = 3). The main effects of LCN2 (F = 15.842, p = 0.004) was significant; HS024 (F = 3.303, p = 0.107) was not. E Quantitative analysis of phosphorylated P65 (P-P65) protein expression (normalized to total P65) ( n = 3). The main effect of HS024 (F = 11.158, p = 0.010) was significant; LCN2 (F = 3.929, p = 0.083) was not. F Quantitative analysis of C1q protein expression (normalized to GAPDH) ( n = 3). The main effects of LCN2 (F = 14.909, p = 0.005) was significant; HS024 (F = 1.731, p = 0.225) was not. G Immunofluorescence analysis after conditional treatment. Representative images of P65 immunoreactivity (green) and DAPI nuclear staining (blue) in BV-2 cells. White arrows indicate P65 nuclear expression-positive cells. H Quantitative analysis of the results in G, ( n = 4). Scale bar: 50 μm. The main effects of LCN2 (F = 12.917, p = 0.004) was significant; HS024 (F = 2.696, p = 0.127) was not. I ELISA measurement of C1q in cell culture supernatant ( n = 4). The main effects of LCN2 (F = 37.783, p < 0.001) and HS024 (F = 9.739, p = 0.009) were both significant. J Western blot analysis to assess the expression levels of TNF-α and TGF-β in four groups. K Quantitative analysis of TNF-α protein expression (normalized to GAPDH) ( n = 3). The main effects of LCN2 (F = 22.812, p = 0.001) and HS024 (F = 96.627, p < 0.001) were both significant. L Quantitative analysis of TGF-β protein expression (normalized to GAPDH) ( n = 3). The main effect of HS024 (F = 65.778, p < 0.001) was significant; LCN2 (F = 0.994, p = 0.348) was not. Data are presented as mean ± SEM. For panels ( A , C - F , H - I , K - L ), data were analyzed by two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns = not significant

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: PF ameliorates neuronal loss induced by silica exposure, which is accompanied by anxiety- and depression-like behaviors in the hippocampus. A Representative activity traces of mice in the OFT across four groups. B Total distance traveled by mice in the OFT ( n = 8). The main effects of Sil (F = 20.072, p < 0.001) and PF (F = 0.637, p = 0.432). C Immobility time in the OFT ( n = 8). The main effects of Sil (F = 20.513, p < 0.001) and PF (F = 5.440, p = 0.027). D Time spent in the central area of the OFT ( n = 8). The main effects of Sil (F = < 0.001, p = 0.983) and PF (F = 0.527, p = 0.474). E Activity trajectories of mice in the open arms and closed arms of the EPM. The red arrow indicates open arms; the green arrow indicates closed arms. F Total distance traveled by mice in the EPM ( n = 8). The main effects of Sil (F = 1.024, p = 0.320) and PF (F = 2.498, p = 0.125). G Time spent in the open arms ( n = 8). The main effects of Sil (F = 1.157, p = 0.291) and PF (F = 5.258, p = 0.030). H Number of entries into the open arms ( n = 8). The main effects of Sil (F = 8.321, p = 0.007) and PF (F = 0.771, p = 0.387). I Number of marbles buried by mice in the MBT ( n = 8). The main effects of Sil (F = 6.215, p = 0.019) and PF (F = 0.559, p = 0.461). J Western blot analysis assessing the expression levels of LCN2, MC4R, C1q, and BDNF in the four groups of mice. K Quantitative analysis of LCN2 protein expression ( n = 9). The main effects of Sil (F = 2.801, p = 0.104) and PF (F = 8.961, p = 0.005). L Quantitative analysis of MC4R protein expression ( n = 9). The main effects of Sil (F = 22.590, p < 0.001) and PF (F = 9.298, p = 0.005). M Quantitative analysis of C1q protein expression ( n = 9). The main effects of Sil (F = 5.380, p = 0.027) and PF (F = 65.328, p < 0.001). N Quantitative analysis of BDNF protein expression ( n = 9). The main effects of Sil (F = 18.296, p < 0.001) and PF (F = 24.246, p < 0.001). Data are presented as mean ± SEM. For panels (B-D, F-I, K-N), data were analyzed by two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns = not significant

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques: Activity Assay, Western Blot, Expressing

Journal: Journal of Neuroinflammation

Article Title: A microglial LCN2-MC4R signaling axis drives silica-induced neuronal damage via C1q release

doi: 10.1186/s12974-026-03695-5

Figure Lengend Snippet: Proposed mechanism by which the microglial LCN2-MC4R signaling axis drives silica-induced neuronal loss. Silica exposure in the lungs triggers the release of LCN2. Both LCN2 and TNF-α then travel to the hippocampus, cross the blood-brain barrier, and activate microglia. In microglia, LCN2 binds to the MC4R, activating the cAMP/PKA/NF-κB signaling pathway. This activation stimulates microglia to release C1q, which initiates the complement cascade, leading to C3 deposition on neurons. These C3-tagged neurons are subsequently targeted by microglia, resulting in neuronal loss

Article Snippet: Cells were cultured in medium containing LCN2 (Sino Biological, 50409-M08H) at 100 ng/ml, the MC4R antagonist HS024 (MedChem Express, HY-P1215A) at 100 nM, or a combination of LCN2 and HS024 (LCN2-HS024) for 24 h. Cell supernatants were collected for subsequent ELISA analysis.

Techniques: Activation Assay

Journal: Cell and Tissue Research

Article Title: Early intestinal barrier changes in A53T transgenic Parkinson’s disease mice

doi: 10.1007/s00441-026-04062-9

Figure Lengend Snippet: Assessment of systemic and local inflammatory markers. a No significant difference in CRP plasma levels was found when comparing between genotypes at either 12 or 36 weeks; however, both WT and A53T mice demonstrated significant age-related reductions. b No significant differences in plasma TNF-α levels were detected. c No significant differences in plasma IL-6 levels were detected; however, an age-related increase was seen in A53T mice. d Plasma LBP levels remained comparable between WT and A53T mice at both 12 and 36 weeks. e , f GFAP-immunoreactivity in the myenteric plexus of the ileum showed no genotype-dependent differences in the distal ileum ( e ) or the distal colon ( f ). g , h CD45-immunoreactivity remained comparable between genotypes in both the ileum ( g ) and colon ( h ). i Representative images of GFAP (cyan) and CD-45 (magenta) in the ileum of 36-week-old A53T mice. Statistical analysis was performed using two-way ANOVA followed by Fisher’s LSD test for genotype comparisons. Results are presented as mean ± SEM, with n = 5–8 per group

Article Snippet: Plasma samples were diluted 1:10 in plasma diluent, and TNF and C-reactive protein (CRP) levels were quantified using mouse TNF (ab208348, Abcam) and CRP (DY1829, R&D systems) ELISA kits following the manufacturer’s protocol.

Techniques: Clinical Proteomics

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin mitigates 2% DSS-induced damage in NCM460 cells. ( A ) Effects of different concentrations of DSS on the viability of NCM460 cells. ( B ) Effects of different concentrations of cordycepin (COR) on the viability of NCM460 cells. NCM460 cells were treated with 2% DSS and / or 1 μmol/L (COR-1), 10 μmol/L (COR-10) COR for 24 h, the release levels of ( C ) LDH, ( D ) IL-1β, ( E ) IL-6, and ( F ) TNF-α in the supernatant were measured by using Elisa. ( G ) Representative images of PI staining and the percentage of PI-positive cells, scar bar = 100 μm. ( H ) The protein expression of ZO-1 in NCM460 cells was determined by using Western blotting assay. The results were representative of three independent experiments and expressed as mean ± SEM. Data were compared using two-tailed Student’s t tests. # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 vs. control group; *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 vs. the 2% DSS group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: Cordycepin abates DSS-induced damage in NCM460 cells by activating adenosine receptor A 2A . ( A ) The cAMP levels in NCM460 cells under treatment with 2% DSS and/or 1 μmol/L and 10 μmol/L COR. ( B ) The cAMP levels in NCM460 cells under treatment with 2% DSS, 10 μmol/L COR, 5 μmol/L SCH58261 (SCH), and / or 5 μmol/L DPCPX for 24 h. The release levels of ( C ) LDH, ( D ) IL-1β, ( E ) IL-6, and ( F ) TNF-α were measured by using Elisa. ( G ) Representative images of PI staining and the percentage of PI-positive cells, scar bar = 100 μm. ( H – J ) The protein expression of ZO-1, A 2A AR and A 1 AR in NCM460 cells was determined by using Western blotting assay. The results were representative of three independent experiments and expressed as mean ± SEM. Data were compared using two-tailed Student’s t tests. vs. control group; # P < 0.05, vs. control group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. the 2% DSS group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Expressing, Western Blot, Two Tailed Test, Control

Journal: Drug Design, Development and Therapy

Article Title: Cordycepin Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice by Inhibiting IL-6/IL-6R-Mediated p38 MAPK and NF-κB Activation Through Adenosine A 2A Receptor Stimulation

doi: 10.2147/DDDT.S575035

Figure Lengend Snippet: SCH58261 blocks cordycepin’s amelioration of intestinal inflammation and gut barrier function in DSS-induced colitis mice. ( A – C ) Level of colonic IL-1β, IL-6, and TNF-α among the Control, DSS, DSS+COR-L and DSS+COR-H group were detected by using Elisa. ( D ) Representative images of AB-PAS staining of colonic tissue. The arrow indicates the goblet cells. ( E ) Number of goblet cells in colonic tissue. ( F ) Representative images of TUNEL staining in colonic tissue, scale bar = 50 μm. ( G ) Percentage of TUNEL-positive cells (%). ( H ) The protein expression of ZO-1 and Occludin in colons was determined by using Western blotting assay. ( I ) and ( J ) Serum level of D-lactate and diamine oxidase (DAO) were measured by using Elisa. ( K ) The serum cAMP levels among the control, DSS, DSS+COR-L, DSS+COR-H group and DSS+SCH+COR-H group. Data were presented as the means ± SEM of six-eight mice in each group and were compared using two-tailed Student’s t tests. # P < 0.05, ## P < 0.01, ### P < 0.001 and #### P < 0.0001 vs. control group; *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 vs. the DSS model (DSS) group.

Article Snippet: Serum CRP levels were measured using a mouse-specific sandwich ELISA kit (E-EL-M0053, Elabscience).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Staining, TUNEL Assay, Expressing, Western Blot, Two Tailed Test

Journal: bioRxiv

Article Title: GP130 Y814 SIGNALING IS REQUIRED FOR THE DYNAMIN-MEDIATED ENDOCYTOSIS, MAPK/P38 ACTIVATION AND PERSISTENCE OF CHRONIC SYSTEMIC INFLAMMATION INDUCED BY HIGH FAT DIET

doi: 10.64898/2026.02.06.704505

Figure Lengend Snippet: (A) Schematic of experimental design. (B) Weight change over time of mice on high fat diet (HFD), n=10 biological replicates per group. (C) Representative images and body composition analysis of WT mice on normal diet (ND) or HFD, generated by DEXA scan. (D) Serum levels of IL-6, CRP, and MCP-1 measured by ELISA in mice fed ND and HFD for 42 weeks. (E) Circulating lymphocytes, neutrophils, and monocytes in peripheral blood from mice on HFD as detected by white blood cell differential. Data points represent biological replicates, error bars are shown as mean +/- SEM, and statistical analyses were performed with 2-way ANOVA with Fisher’s LSD test for more than two groups or Student’s t-test for comparing two groups; p-values less than 0.05 were considered significant.

Article Snippet: ELISA kits (mouse CRP (MCRP00), mouse IL-6 (M6000B), mouse CCL2/MCP-1 (MJE00B)) were purchased from R&D Systems and performed following the manufacturer’s instruction.

Techniques: Generated, Enzyme-linked Immunosorbent Assay