elisa kit  (Revvity)

Journal: Gut Microbes

Article Title: Clostridioides difficile aggravates dextran sulfate solution (DSS)-induced colitis by shaping the gut microbiota and promoting neutrophil recruitment

doi: 10.1080/19490976.2023.2192478

Figure Lengend Snippet: The gut microbiota is altered in mice with colitis challenged with C. difficile . (a – d) the effect of C. difficile on mice with colitis was independent of toxin production, as indicated by weight loss (a), H&E staining (b), histological scores (c), and production of IL-6 as shown by ELISA (d). Scale bar, 200 μm. (e and f) PcoA at the OTU level for samples from feces (e) and colonic contents (f), and the plots were based on the Bray – Curtis distance. The horizontal and vertical axes represent inter-sample variations. Each point represents an individual sample, and different colors refer to different groups. (g) Cladograms were generated by linear discriminant analysis effect size analysis to detect the differences in bacterial taxa between the DSSCD and DSS groups. Circles show phylogenetic levels from the phylum to the genus. To screen out differentially abundant taxa, the linear discriminant analysis threshold score was set to>4.0. Red and blue bars indicate taxa enrichment in the DPI48H_DSS and DPI48H_DSSCD groups, respectively. (h) Correlations between IL-6 levels and relative abundance of g_prevotellaceae_ucg001 (OTU288) in DSSCD groups (DPI6H_DSSCD and DPI48_DSSCD) were analyzed by Spearman’s correlation. Each dot represents a value from an individual mouse. Data are expressed as the mean±sd. Statistical differences between groups were assessed by the Mann – Whitney test. * P < 0.05.

Article Snippet: The production of IL-6 was also measured by the enzyme-linked immunosorbent assay (ELISA) kit (Biolegend, USA).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Generated, MANN-WHITNEY

Journal: Gut Microbes

Article Title: Clostridioides difficile aggravates dextran sulfate solution (DSS)-induced colitis by shaping the gut microbiota and promoting neutrophil recruitment

doi: 10.1080/19490976.2023.2192478

Figure Lengend Snippet: Altered gut microbiota in DSSCD group contributes to the severity of colitis. (a) Experimental design of FMT. The mice were pretreated with an antibiotic cocktail for 3 weeks, followed by 2% DSS treatment and oral gavage of fecal samples derived from DSSCD and DSS group every other day during modeling. (b – e) the severity of colitis in the mouse transplanted fecal microbiota in the DSSCD and DSS groups was assessed by body weight loss (b), H&E and alcian blue staining (c), histological scores (d), and IL-6 levels from colonic tissue (e). Scale bars, 200 μm. (f) the relative abundance of OTU288 and OTU287 was quantified by real-time qPCR with normalization to total bacterial (16S rRNA). (g – h) the effect of C. difficile on mice with colitis and antibiotic pretreatment was evaluated by weight loss (g), histological scores from H&E staining (h), and colonic IL-6 production by ELISA (i). Scale bars, 200 μm. Data are shown as the mean±sd and represent at least three independent experiments. Statistical analysis between groups was conducted by the Mann – Whitney test. * P < 0.05, ** P < 0.01.

Article Snippet: The production of IL-6 was also measured by the enzyme-linked immunosorbent assay (ELISA) kit (Biolegend, USA).

Techniques: Derivative Assay, Staining, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Molecules

Article Title: Beneficial Effects of Hordenine on a Model of Ulcerative Colitis

doi: 10.3390/molecules28062834

Figure Lengend Snippet: Hordenine ameliorated DSS-induced UC in mice. ( A ) The chemical structure of hordenine. ( B ) Flow chart of the experimental design. SPSS, as known as stroke-physiological saline solution; ELISA: enzyme linked immunosorbent; HE: hematoxylin-eosin staining; IHC: immunohistochemistry; WB: western blot; PAS: periodic acid-Schiff (PAS) staining; the elements in the graph are derived from figdrow. ( C ) Changes in body weight. ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. DSS group. ( D ) The DAI scores were calculated. ( E ) The macroscopic representations of colons. ( F ) The colon lengths among the six groups. ** p < 0.01 vs. control group; # p < 0.01 vs. DSS group. ( G ) Hematoxylin and eosin staining of colonic sections. Ulcer and necrosis are indicated by black arrow. Inflammatory cell infiltration and aggregation are indicated by red arrow. Tissue edema is indicated by blue arrow. Scale bar: 100 μm. ( H ) Histological changes. ** p < 0.05 compared with control group; ## p < 0.05 compared with DSS group. ( I ) periodic acid-Schiff staining of colonic sections. ( J ) Goblet cells count. ** p < 0.05 compared with control group; ## p < 0.05 compared with DSS group.

Article Snippet: Mouse interleukin (IL)-6, IL-1β, and tumor necrosis factor-alpha (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Biolegend (San Diego, CA, USA).

Techniques: Saline, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Western Blot, Derivative Assay, Control

Journal: Molecules

Article Title: Beneficial Effects of Hordenine on a Model of Ulcerative Colitis

doi: 10.3390/molecules28062834

Figure Lengend Snippet: Hordenine (50 mg/kg) reduces levels of inflammatory mediators in the colon of mice with dextran sodium sulfate (DSS)-induced colitis. Enzyme-linked immunosorbent analysis of IL-6 ( A ), IL-1β ( B ) and TNF-α ( C ) in colon tissue supernatants. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. DSS group. ( D ) Immunohistochemical staining of IL-6 and TNF-α in colon tissues.Red arrows point to the positive expression of the target protein, scale bar: 50 μm. ( E – H ) Western blotting analysis of IL-6, IL-1β and TNF-α expression in colon tissues. * p < 0.05, ** p < 0.01 vs. control group; # p < 0.05, ## p < 0.01 vs. DSS group.

Article Snippet: Mouse interleukin (IL)-6, IL-1β, and tumor necrosis factor-alpha (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Biolegend (San Diego, CA, USA).

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

Journal: The Journal of Clinical Investigation

Article Title: Clonally expanded HIV-1 proviruses with 5 ′ -leader defects can give rise to nonsuppressible residual viremia

doi: 10.1172/JCI165245

Figure Lengend Snippet: ( A ) Deletions found in proviruses causing viremia were introduced in an NL4-3 expression plasmid by site-directed mutagenesis. Deletion start and end positions relative to HXB2 are indicated in parentheses. ( B ) Copies of HIV-1 RNA recovered at 72 hours after transfection of 293T cells, expressed as copies per mL (left) or normalized by p24 pg/mL (right). ( C ) Spinoculation of primary CD4 + T cells shows exponential increase in p24 levels only with WT NL4-3, in the absence of antiretrovirals (ARVs: TDF, FTC, DTG). ( D ) Reverse transcription was assessed by measuring late cDNA products by ddPCR targeting the U5-PBS junction. Primary CD4 + T cells were collected at 0, 6, and 12 hours after spinoculation with and without ARVs. U5-PBS copies detected in the presence of ARVs, which are the result of incomplete DNAseI digestion of plasmid carryover from transfection, were subtracted from copies detected in conditions without ARVs. ( E ) Virus produced upon 293T transfection was pelleted by ultracentrifugation, lysed, normalized by p24, and used for Western blots with primary antibodies specific to p24 and gp41. ( F ) Surface staining of HIV-1 env on 293T cells 24 hours after transfection. ( G ) Frequency of env-positive cells transfected with WT versus 5 ′ -L deletions. Fold reduction relative to WT is indicated above each mutant. Results from 2 transfection experiments are shown. Each circle represents the average of 2 technical replicates. ( H ) Quantification of cell-associated spliced HIV-1 transcripts belonging to the 4 kb class or tat/rev mRNA normalized to RNA ng. ( I ) Percentage of spliced transcripts relative to total HIV-1 polyA RNA. Error bars indicate SEM ( B , H and I ) or SD ( C and F ). Statistical significance between conditions was determined by 1-way ANOVA. * P < 0.05; ** P <0.01, *** P <0.001.

Article Snippet: Virus recovery was measured by p24 ELISA (PerkinElmer) and reverse transcriptase PCR (RT-PCR) measuring polyadenylated HIV-1 RNA.

Techniques: Expressing, Plasmid Preparation, Mutagenesis, Transfection, Reverse Transcription, Virus, Produced, Western Blot, Staining