elisa kits  (R&D Systems)

Journal: Scientific Reports

Article Title: Plant extract mixture shows anti-inflammatory and barrier-strengthening effects and activates aryl hydrocarbon receptor in a 2D psoriasis model

doi: 10.1038/s41598-026-50000-8

Figure Lengend Snippet: Plant extract mixture downregulates inflammatory markers in the 2D psoriasis model. CaCl 2 -differentiated NHEKs were stimulated as described in Fig. . Gene expression levels of ( a ) IL1A , ( b ) IL1B, ( c ) IL1RN , ( d ) CXCL8 , ( e ) TNFA , ( f ) IL17C , ( g ) IL36G , ( h ) CSF2 , ( i ) VEGFA were measured. Statistical significance was tested by a, b, c, e, h) one-way ANOVA with subsequent Sidak’s multiple comparison test or d, f, g) Kruskal-Wallis test with subsequent Dunn’s multiple comparisons test ( n = 12, * p < 0.05; ** p < 0.01; *** p < 0.001; ns = not significant). Protein expression levels of ( j ) IL-8 and k) TNFα were measured in the supernatants of the cells by ELISA. Values for unstimulated control and plant extract-treated cells were below the detection limit of 31,3 pg/mL (IL-8) or 62,5 pg/mL (TNFα) and not detectable; therefore, no statistical analysis was performed. n.d.= non-detectable.

Article Snippet: TNFα and IL-8 concentrations were determined using commercial ELISA kits (human TNFα or human CXCL8/IL-8 DuoSet ELISA, R&D Systems, Minneapolis, MN, USA) following the manufacturer’s protocol.

Techniques: Plant Extract, Gene Expression, Comparison, Expressing, Enzyme-linked Immunosorbent Assay, Control

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: Infection and Immunity

Article Title: Loss of Fgr41 in Candida albicans attenuates virulence and increases proinflammatory immune responses in a manner that is dependent on β(1,3)-glucan but not dectin-1

doi: 10.1128/iai.00523-25

Figure Lengend Snippet: FGR41 knockout-induced β(1, 3)-glucan exposure and macrophage TNF-α release are decreased by ENG1 overexpression. ( A ) Cells were stained for confocal microscopy with a primary antibody to β(1, 3)-glucan and a secondary antibody conjugated to Alexa Fluor 488 to visualize unmasking, and CFW to visualize total chitin. A total of 30 images was taken of each strain across three biological replicates. Scale bar = 8 µm. ( B ) Quantification of the exposed β(1, 3)-glucan in panel A. A global threshold of 60 was applied to each exposed β(1, 3)-glucan image, and the percentage of cells showing fluorescence in each image was determined using Fiji ImageJ. **** P < 0.0001, ** P < 0.01, * P < 0.05, by one-way analysis of variance (ANOVA). ( C ) The wild type and fgr41Δ/Δ bars from panel B were analyzed in isolation, which revealed a significant difference (** P < 0.01 by one-way ANOVA). ( D ) RAW 264.7 macrophages were co-incubated with UV-inactivated C. albicans for 4 h, and the resulting supernatants were analyzed for TNF-α concentrations via ELISA. The experiment was performed three times with three technical replicates each time; each point represents the average of three technical replicates. **** P < 0.0001, ** P < 0.01, * P < 0.05, by one-way ANOVA, ns = not significant.

Article Snippet: Mouse TNF-α ELISA kits were purchased from R&D Systems (DY410).

Techniques: Knock-Out, Over Expression, Staining, Confocal Microscopy, Fluorescence, Isolation, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Loss of Fgr41 in Candida albicans attenuates virulence and increases proinflammatory immune responses in a manner that is dependent on β(1,3)-glucan but not dectin-1

doi: 10.1128/iai.00523-25

Figure Lengend Snippet: Effect of a neutralizing dectin-1 antibody or inhibitory laminarin on TNF-α release from macrophages stimulated by eng1Δ/Δ and fgr41Δ/Δ mutants. RAW 264.7 macrophages were treated with dectin-1 antibody (100 ng/mL) or inhibitory laminarin (100 µg/mL) for 1 h prior to a 4 h co-incubation with UV-inactivated C. albicans strains. TNF-α release was measured using an ELISA. Each experiment was performed four times with three replicates each time; each point represents the average of three technical replicates. ( A ) Unchallenged macrophages show no differences in TNF-α release when pretreated with dectin-1 antibody or laminarin. ns = not significant. ( B ) Wild type-challenged macrophages show a small decrease in TNF-α release after both dectin-1 antibody and laminarin pretreatment. ** P < 0.01, * P < 0.05, by one-way ANOVA, ns = not significant. ( C ) eng1Δ/Δ -challenged macrophages exhibit a small but insignificant decrease in TNF-α release after pretreatment with dectin-1 antibody, and a significant decrease after pretreatment with laminarin. *** P < 0.001, * P < 0.05, by one-way ANOVA, ns = not significant. ( D ) Pretreatment with dectin-1 antibody did not significantly impact TNF-α release from macrophages incubated with fgr41Δ/Δ, although a small decrease was observed. Pretreatment with laminarin caused a large, significant reduction. * P < 0.05 by one-way ANOVA, ns = not significant.

Article Snippet: Mouse TNF-α ELISA kits were purchased from R&D Systems (DY410).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Infection and Immunity

Article Title: Loss of Fgr41 in Candida albicans attenuates virulence and increases proinflammatory immune responses in a manner that is dependent on β(1,3)-glucan but not dectin-1

doi: 10.1128/iai.00523-25

Figure Lengend Snippet: Proposed model. Eng1 localizes to bud necks and scars and degrades β(1, 3)-glucan at the septum during cell division. Eng1 may also be interacting with Fgr41 at the cell surface and can partially compensate for the loss of Fgr41 in unmasking and TNF-α stimulation. While exposed β(1, 3)-glucan in a wild-type cell is recognized primarily by dectin-1 on the surface of macrophages, recognition of the fgr41Δ/Δ mutant appears to be dectin-1-independent and is likely driven primarily by other receptors. Once the macrophage has recognized the fgr41Δ/Δ mutant, levels of phosphorylated p38 increase, and by this or other components, the signal is transduced to a transcription factor other than NF-κB to lead to TNF-α production.

Article Snippet: Mouse TNF-α ELISA kits were purchased from R&D Systems (DY410).

Techniques: Mutagenesis