immunosorbent assay elisa max standard kit  (Revvity)

Journal: International Journal of Molecular Sciences

Article Title: TLR9 Signaling Is Required for the Porphyromonas gingivalis -Induced Activation of IL-10-Expressing B Cells

doi: 10.3390/ijms24076693

Figure Lengend Snippet: Under P. gingivalis stimulation for 24 h, the CD45 + IL-10 + cells, IL-10 mRNA level and secreted IL-10 protein levels of splenocytes and B cells isolated from pre-immunized TLR9 knockout mice were calculated. ( A ) The representative ICC images of CD45 + IL-10 + cells in each group. CD45 cells were labeled with rat anti-mouse CD45 antibody conjugated with emerald anti-rat antibody (blue) and IL-10 were labeled with goat anti-mouse IL-10 antibody conjugated with permanent red anti-goat antibody (red). ( B , E ) The percentage of CD45 + IL-10 + cells among splenocytes ( B ) and B cells ( E ) were calculated based on cell immunocytochemistry staining. ( C , F ) The IL-10 mRNA expression in splenocytes ( C ) and B cells ( F ) was analyzed by quantitative RT-PCR. ( D , G ) The secreted IL-10 protein levels in the supernatants were measured by a mouse IL-10 enzyme-linked ELISA kit using a 1:10 assay diluent in PBST. The data were shown as mean ± SD; significance calculated by unpaired t -test was indicated as ** p < 0.01, **** p < 0.0001, and ns (no significant difference) as p > 0.05 (n = 5).

Article Snippet: The secreted IL-10 protein levels in the supernatants of cultured splenocytes and B cells were measured by a mouse IL-10 enzyme-linked immunosorbent assay (ELISA) Max Standard kit (BioLegend, San Diego, CA, USA) using 1:10 assay diluent in PBST.

Techniques: Isolation, Knock-Out, Labeling, Immunocytochemistry, Staining, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: TLR9 Signaling Is Required for the Porphyromonas gingivalis -Induced Activation of IL-10-Expressing B Cells

doi: 10.3390/ijms24076693

Figure Lengend Snippet: Under P. gingivalis stimulation for 24 h, the CD45 + IL-10 + cells of splenocytes and B cells, and secreted IL-10 protein levels of B cells isolated from both pre-immunized wild-type and TLR9 knockout mice were calculated. ( A , B ) The percentage of CD45 + IL-10 + cells among splenocytes ( A ) and B cells ( B ) were calculated based on cell immunocytochemistry staining. ( C ) The secreted IL-10 protein levels in the supernatants were measured by a mouse IL-10 enzyme-linked ELISA kit using a 1:10 assay diluent in PBST. The data were shown as mean ± SD; significance calculated by unpaired t -test was indicated as * p < 0.05, **** p < 0.0001 (n = 5).

Article Snippet: The secreted IL-10 protein levels in the supernatants of cultured splenocytes and B cells were measured by a mouse IL-10 enzyme-linked immunosorbent assay (ELISA) Max Standard kit (BioLegend, San Diego, CA, USA) using 1:10 assay diluent in PBST.

Techniques: Isolation, Knock-Out, Immunocytochemistry, Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Personalized neoantigen viro-immunotherapy platform for triple-negative breast cancer

doi: 10.1136/jitc-2023-007336

Figure Lengend Snippet: Validation of candidate MDA-MB-231-derived neoantigens. (A) For determination of cytotoxic activity of neoantigen-stimulated CD8+T cells, the bulk of CD8+T cells from donors 115 and 118 stimulated with the indicated neoantigen peptides were incubated overnight with carboxyfluorescein succinimidyl ester (CFSE)-labeled T2 cells pulsed with the neoantigen (red) or CFSE-labeled MDA-MB-231 cells (blue). Cell viability of target T-cells in these co-cultures was determined by staining with EthD-1 followed by flow cytometry analysis. Target T-cells gated on the CFSE-positive population that were positive for EthD-1 were considered dead cells. (B, C) CD8+T cells from a HLA-A*02:01-positive donor were stimulated as in (A) with peptide #22 and incubated overnight with peptide #22-negative/HLA-A*02:01-positive BT549 or peptide #22-positive/HLA-A*02:01-positive MDA-MB-231 cells with or without blocking anti-MHC-I antibodies. (B) Supernatants were collected and assayed for IFN-γ by ELISA and (B) cytotoxicity was determined as in (A). The data were analyzed by one-way ANOVA*p<0.05; **p<0.01; ***p<0.001. Graph shows mean±SEM. ANOVA, analysis of variance.

Article Snippet: IFN-γ levels from the co-cultures were measured by ELISA Max Standard kit (BioLegend) in 96-well microtiter plates according to the manufacturer’s instructions.

Techniques: Biomarker Discovery, Derivative Assay, Immunopeptidomics, Activity Assay, Incubation, Labeling, Staining, Flow Cytometry, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: eBioMedicine

Article Title: Dysregulation of the kallikrein-kinin system in bronchoalveolar lavage fluid of patients with severe COVID-19

doi: 10.1016/j.ebiom.2022.104195

Figure Lengend Snippet: MPO-DNA complexes in bronchoalveolar lavage fluid from patients with and without COVID-19 . Graphs show individual dots representing patient data and bar representing median of each patient group. For graphical representation, data were transformed using the logarithmic transformation. Pairwise comparisons were made between patients with COVID-19 ( n = 21) and patients without COVID-19 ( n = 19). Median [range] and p -value are shown (Mann-Whitney U test).

Article Snippet: A standard curve was prepared by incubating a known quantity of an in-house-generated MPO-DNA mixture complex consisting of enzymatically inactive MPO standard (from the LEGEND MAX Human MPO ELISA kit, Biolegend), lambda-DNA (Invitrogen) and human native nucleosomes (Sigma).

Techniques: Transformation Assay, MANN-WHITNEY

Journal: Arthritis Research & Therapy

Article Title: Semaphorin 3G exacerbates joint inflammation through the accumulation and proliferation of macrophages in the synovium

doi: 10.1186/s13075-022-02817-7

Figure Lengend Snippet: Attenuated joint inflammation in Sema3G-deficient mice. A The clinical score of Sema3G-deficient (Sema3G −/− ) mice and their littermate controls (Sema3G +/− ) in CIA. Data were obtained from four independent experiments ( N = 17 in each group). B The pathological scores of CIA. The front paws were subjected to H&E staining, and the inflammation, the cartilage damage, and the erosion scores were separately assessed. C The inflammatory cytokines in sera. The sera were collected on day 42 and subjected to ELISA to measure IL-6 and TNFα. D The clinical score of Sema3G −/− mice and Sema3G +/− in CAIA. Data were obtained from four independent experiments ( N = 13 in each group). E The pathological scores of CAIA. The front paws were subjected to H&E staining, and the inflammation, the cartilage damage, and the erosion scores were separately assessed. F The inflammatory cytokines in sera. The sera were collected on day 9 and subjected to ELISA to measure IL-6 and TNFα. Data are expressed as the means ± SEM. For the analyses of the clinical score, 2-way ANOVA was used. For the pathological scores, an unpaired t -test was used

Article Snippet: The serum levels of IL-6 and TNFα were measured using a mouse ELISA MAX Standard IL-6 kit (BioLegend) and a mouse ELISA MAX Standard TNFα kit (BioLegend).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Arthritis Research & Therapy

Article Title: Semaphorin 3G exacerbates joint inflammation through the accumulation and proliferation of macrophages in the synovium

doi: 10.1186/s13075-022-02817-7

Figure Lengend Snippet: Attenuated joint inflammation in Sema3G-deficient mice. A The clinical score of Sema3G-deficient (Sema3G −/− ) mice and their littermate controls (Sema3G +/− ) in CIA. Data were obtained from four independent experiments ( N = 17 in each group). B The pathological scores of CIA. The front paws were subjected to H&E staining, and the inflammation, the cartilage damage, and the erosion scores were separately assessed. C The inflammatory cytokines in sera. The sera were collected on day 42 and subjected to ELISA to measure IL-6 and TNFα. D The clinical score of Sema3G −/− mice and Sema3G +/− in CAIA. Data were obtained from four independent experiments ( N = 13 in each group). E The pathological scores of CAIA. The front paws were subjected to H&E staining, and the inflammation, the cartilage damage, and the erosion scores were separately assessed. F The inflammatory cytokines in sera. The sera were collected on day 9 and subjected to ELISA to measure IL-6 and TNFα. Data are expressed as the means ± SEM. For the analyses of the clinical score, 2-way ANOVA was used. For the pathological scores, an unpaired t -test was used

Article Snippet: The serum levels of IL-6 and TNFα were measured using a mouse ELISA MAX Standard IL-6 kit (BioLegend) and a mouse ELISA MAX Standard TNFα kit (BioLegend).

Techniques: Staining, Enzyme-linked Immunosorbent Assay