igg quantitation enzyme linked immunosorbent assay  (Bethyl)

 
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    Structured Review

    Bethyl igg quantitation enzyme linked immunosorbent assay
    Igg Quantitation Enzyme Linked Immunosorbent Assay, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg quantitation enzyme linked immunosorbent assay/product/Bethyl
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    igg quantitation enzyme linked immunosorbent assay - by Bioz Stars, 2021-07
    93/100 stars

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    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Metformin rescues rapamycin-induced mitochondrial dysfunction and attenuates rheumatoid arthritis with metabolic syndrome
    Article Snippet: Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. .. Measurement of immunoglobulin G concentrations The immunoglobulin G (IgG) concentrations were measured using mouse IgG ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX, USA) in splenocyte culture supernatant. .. Flow cytometry Before intracellular staining, cells were stimulated with 25 ng/ml phosphomolybdic acid (Sigma-Aldrich), 250 ng/ml ionomycin (Sigma-Aldrich), and Golgi Stop (BD Biosciences, San Diego, CA, USA) in 5% CO2 at 37 °C for 4 h. Cells were stained with peridinin chlorophyll protein complex–conjugated anti-CD4 and allophycocyanin (APC)-conjugated anti-CD25 antibodies (BD Pharmingen, BD Biosciences), and then with a phycoerythrin (PE)-conjugated anti-Foxp3 antibody (eBioscience, San Diego, CA, USA), followed by fixation and permeabilization using a Cytofix/Cytoperm Plus Kit (BD Biosciences) according to the manufacturer’s instructions.

    Article Title: Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease
    Article Snippet: The results of the reactions were reported as positive or negative, and the pattern of fluorescence was described as advised for IgG antinuclear antibodies. .. Enzyme-linked immunosorbent assay (ELISA) The concentration of IgE, IgA and IgG was determined by ELISA in the serum of SLE patients using the human ELISA quantitation set (Bethyl Laboratories, Inc, Montgomery, TX, USA) according to the manufacturer’s instructions. .. Individual sera diluted at 1:4, 1:50 000 and 1:100 000 for IgE, IgA and IgG respectively were subjected to ELISA analysis, and concentrations in individual samples were calculated according to the standard curve.

    Article Title: The comparative immunotoxicity of mesoporous silica nanoparticles and colloidal silica nanoparticles in mice
    Article Snippet: Antibodies to cell-specific receptors (Sasnta Cruz Biotechnology, Santa Cruz, CA, USA) included hamster anti-mouse CD3- FITC (T lymphocyte), rat anti-mouse CD45R/B220-PE (B lymphocyte), rat anti-mouse CD4-FITC (T helper lymphocyte), and rat anti-mouse CD8-PE (T cytotoxic/suppressor lymphocyte). .. Measurement of serum immunoglobulin G (IgG) and immunoglobulin M (IgM) Immunoglobulin levels were analyzed using a mouse IgG and IgM enzyme-linked immunsorbent assay (ELISA) Quantitation Kit (Bethyl Laboratories, Montgomery, TX, USA) following the manufacturer’s instructions. .. Briefly, 96-well plates were coated with affinity purified mouse IgG and IgM coating antibody.

    Article Title: Serum Antibody Response to Three Non-typeable Haemophilus influenzae Outer Membrane Proteins During Acute Otitis Media and Nasopharyngeal Colonization in Otitis Prone and Non-Otitis Prone Children
    Article Snippet: .. The levels of IgG, IgM and IgA in the reference serum were quantitatively measured by using a human IgG/IgA/IgM ELISA quantitation kit (Bethyl laboratories). ..

    Article Title: Metformin Attenuates Experimental Autoimmune Arthritis through Reciprocal Regulation of Th17/Treg Balance and Osteoclastogenesis
    Article Snippet: Stained sections were analyzed using a Zeiss microscope (LSM 510 Meta; Carl Zeiss, Oberkochen, Germany). .. Measurement of Immunoglobulin (Ig) Concentrations Anti-IgG, IgG1, and IgG2a were measured by mouse IgG, IgG1, and IgG2a ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX) [ ]. .. Flow Cytometry of Intracellular Cytokines To analyze intracellular cytokines, splenocytes were stained with PerCP-conjugated anti-CD4, APC-conjugated anti-CD25, FITC-conjugated anti-IL-17, and PE-conjugated anti-Foxp3 (eBiosciences), followed by fixation and permeabilization with a Foxp3 staining buffer kit (BD Bioscience) according to the manufacturer's instructions.

    Article Title: A benzenediamine derivate FC-99 attenuates lupus nephritis in MRL/lpr mice via inhibiting myeloid dendritic cell-secreted BAFF
    Article Snippet: Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St Louis, USA). .. Enzyme-linked immunosorbent assay (ELISA) kits for IgG, IgG2a, and IgM, and Mouse Albumin ELISA Quantitation Set were obtained from Bethyl Laboratories (Montgomery, USA). .. BAFF ELISA kit and antibodies against mouse C3c, BAFF, and IgG were purchased from Abcam (Cambridge, USA).

    Quantitation Assay:

    Article Title: Metformin rescues rapamycin-induced mitochondrial dysfunction and attenuates rheumatoid arthritis with metabolic syndrome
    Article Snippet: Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. .. Measurement of immunoglobulin G concentrations The immunoglobulin G (IgG) concentrations were measured using mouse IgG ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX, USA) in splenocyte culture supernatant. .. Flow cytometry Before intracellular staining, cells were stimulated with 25 ng/ml phosphomolybdic acid (Sigma-Aldrich), 250 ng/ml ionomycin (Sigma-Aldrich), and Golgi Stop (BD Biosciences, San Diego, CA, USA) in 5% CO2 at 37 °C for 4 h. Cells were stained with peridinin chlorophyll protein complex–conjugated anti-CD4 and allophycocyanin (APC)-conjugated anti-CD25 antibodies (BD Pharmingen, BD Biosciences), and then with a phycoerythrin (PE)-conjugated anti-Foxp3 antibody (eBioscience, San Diego, CA, USA), followed by fixation and permeabilization using a Cytofix/Cytoperm Plus Kit (BD Biosciences) according to the manufacturer’s instructions.

    Article Title: Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease
    Article Snippet: The results of the reactions were reported as positive or negative, and the pattern of fluorescence was described as advised for IgG antinuclear antibodies. .. Enzyme-linked immunosorbent assay (ELISA) The concentration of IgE, IgA and IgG was determined by ELISA in the serum of SLE patients using the human ELISA quantitation set (Bethyl Laboratories, Inc, Montgomery, TX, USA) according to the manufacturer’s instructions. .. Individual sera diluted at 1:4, 1:50 000 and 1:100 000 for IgE, IgA and IgG respectively were subjected to ELISA analysis, and concentrations in individual samples were calculated according to the standard curve.

    Article Title: The comparative immunotoxicity of mesoporous silica nanoparticles and colloidal silica nanoparticles in mice
    Article Snippet: Antibodies to cell-specific receptors (Sasnta Cruz Biotechnology, Santa Cruz, CA, USA) included hamster anti-mouse CD3- FITC (T lymphocyte), rat anti-mouse CD45R/B220-PE (B lymphocyte), rat anti-mouse CD4-FITC (T helper lymphocyte), and rat anti-mouse CD8-PE (T cytotoxic/suppressor lymphocyte). .. Measurement of serum immunoglobulin G (IgG) and immunoglobulin M (IgM) Immunoglobulin levels were analyzed using a mouse IgG and IgM enzyme-linked immunsorbent assay (ELISA) Quantitation Kit (Bethyl Laboratories, Montgomery, TX, USA) following the manufacturer’s instructions. .. Briefly, 96-well plates were coated with affinity purified mouse IgG and IgM coating antibody.

    Article Title: Serum Antibody Response to Three Non-typeable Haemophilus influenzae Outer Membrane Proteins During Acute Otitis Media and Nasopharyngeal Colonization in Otitis Prone and Non-Otitis Prone Children
    Article Snippet: .. The levels of IgG, IgM and IgA in the reference serum were quantitatively measured by using a human IgG/IgA/IgM ELISA quantitation kit (Bethyl laboratories). ..

    Article Title: Metformin Attenuates Experimental Autoimmune Arthritis through Reciprocal Regulation of Th17/Treg Balance and Osteoclastogenesis
    Article Snippet: Stained sections were analyzed using a Zeiss microscope (LSM 510 Meta; Carl Zeiss, Oberkochen, Germany). .. Measurement of Immunoglobulin (Ig) Concentrations Anti-IgG, IgG1, and IgG2a were measured by mouse IgG, IgG1, and IgG2a ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX) [ ]. .. Flow Cytometry of Intracellular Cytokines To analyze intracellular cytokines, splenocytes were stained with PerCP-conjugated anti-CD4, APC-conjugated anti-CD25, FITC-conjugated anti-IL-17, and PE-conjugated anti-Foxp3 (eBiosciences), followed by fixation and permeabilization with a Foxp3 staining buffer kit (BD Bioscience) according to the manufacturer's instructions.

    Article Title: A benzenediamine derivate FC-99 attenuates lupus nephritis in MRL/lpr mice via inhibiting myeloid dendritic cell-secreted BAFF
    Article Snippet: Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St Louis, USA). .. Enzyme-linked immunosorbent assay (ELISA) kits for IgG, IgG2a, and IgM, and Mouse Albumin ELISA Quantitation Set were obtained from Bethyl Laboratories (Montgomery, USA). .. BAFF ELISA kit and antibodies against mouse C3c, BAFF, and IgG were purchased from Abcam (Cambridge, USA).

    ChIP-sequencing:

    Article Title: Bcl11b, a novel GATA3-interacting protein, suppresses Th1 while limiting Th2 cell differentiation
    Article Snippet: 220 ng double-stranded DNA was sonicated into 200–400 bp in size using the Bioruptor Pico (Diagenode). .. ChIP-seq experiments were performed with an iDeal ChIP-Seq kit for Transcription Factors (C01010055; Diagenode) using anti-Bcl11b (A300-385A-2; Bethyl Laboratories) and anti-GATA3 (L50-823; BD Biosciences), and iDeal ChIP-seq kit for Histones (C01010051; Diagenode) using anti-H3K27ac (C15410196; Diagenode). .. Both cDNA and ChIP DNA were blunt-ended, ligated to the Solexa adaptors, amplified, and sequenced with an Illumina HiSeq system, and 50-bp reads were generated by the National Heart, Lung, and Blood Institute DNA Sequencing and Computational Biology Core.

    Incubation:

    Article Title: Evaluation of the Efficacy of a Cholera Toxin-Based Staphylococcus aureus Vaccine against Bovine Intramammary Challenge
    Article Snippet: After washing, plates were incubated with two-fold dilutions of either bovine serum (dilutions initiated at 1:200 concentration) or pooled quarter milk (dilutions initiated at a 1:10 concentration). .. After washing, plates were incubated with horseradish peroxidase (HRP)-conjugated anti-bovine IgG, IgG1, IgG2, or IgA (1:10,000 Bethyl Laboratories, Montgomery, TX, USA) at 37 °C for 1 h. Plates were developed with tetramethylbenzidine (PromegaTM TMB One, Thermo Fisher, Waltham, MA, USA) and read at 370 nm per TMB manufacturer’s instruction. .. ELISA results from serum or pooled quarter milk were reported by cow (n = 3) and presented as the ratio of results (day X/day 1) of the O.D. (370 nm) from a representative antibody dilution in the linear part of the curve (1:1600 serum, 1:160 milk).

    Concentration Assay:

    Article Title: Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease
    Article Snippet: The results of the reactions were reported as positive or negative, and the pattern of fluorescence was described as advised for IgG antinuclear antibodies. .. Enzyme-linked immunosorbent assay (ELISA) The concentration of IgE, IgA and IgG was determined by ELISA in the serum of SLE patients using the human ELISA quantitation set (Bethyl Laboratories, Inc, Montgomery, TX, USA) according to the manufacturer’s instructions. .. Individual sera diluted at 1:4, 1:50 000 and 1:100 000 for IgE, IgA and IgG respectively were subjected to ELISA analysis, and concentrations in individual samples were calculated according to the standard curve.

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  • igg2  (Bethyl)
    93
    Bethyl igg2
    Serum IgG1, <t>IgG2,</t> and cytokine expression analysis. ( A ) Anti-IsdA-CTA 2 /B IgG1, ( B ) anti-IsdA-CTA 2 /B IgG2, ( C ) anti-ClfA-CTA 2 /B IgG1, and ( D ) anti-ClfA-CTA 2 /B IgG2 responses in serum on days 14, 20, and 30. Results are reported as ELISA ratios of day X/day 1 at O.D. 370 at serum dilutions of 1:1600. Shown are mean and standard error by treatment with control (gray) and vaccinated (blue) ( n = 3 per group). The log10 of the values were analyzed using analysis of variance (ANOVA), with a compound symmetric covariance structure for cows across days. Model-based estimates were compared between groups within days and adjusted for multipl e comparisons. ( E ) IL-10, IL-6, and IFN-γ expression as determined by quantitative RT-PCR of peripheral blood mononuclear cells (PBMCs) isolated from whole blood after boost on day 20. Results are shown as relative gene expression to GAPDH (2 −ΔΔCt ). Data are presented as mean and standard error of control (gray) and vaccinated (blue) showing median and range ( n = 3 per group). Data were analyzed using a two-group t -test between vaccinated and control. Significant differences between groups are represented as p ≤ 0.05 (*).
    Igg2, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Bethyl immunoglobulin g igg concentrations
    Rapamycin suppresses T cell alloreactivity, proinflammatory cytokines, and production of immunoglobulin and induces mitochondrial dysfunction in vitro. a In a mixed lymphocyte reaction assay, B/c splenic T cells (responders) were incubated with irradiated B/c splenic antigen-presenting cells for 4 days. Responder cells were cultured in the presence or absence of rapamycin. b IFN-γ and IL-17 levels in the supernatants, as measured by enzyme-linked immunosorbent assay (ELISA). c IL-17, IL-6, TNF-α, and <t>IgG</t> concentrations in culture supernatants in the presence of anti-CD3 (0.5 μg/ml) or lipopolysaccharide (100 ng/ml) with rapamycin 100 nM, as determined by ELISA. d The Treg population, as determined by flow cytometry. e JC-1 populations in isolated splenic cells, as determined by flow cytometry. The data are representative of at least three independent experiments. Data represent the mean ± SD of three independent experiments. ** P
    Immunoglobulin G Igg Concentrations, supplied by Bethyl, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Bethyl mouse igg
    Therapeutic effects of metformin in CIA model. CIA was induced in C57BL/6 mice. Metformin 5 mg/mice ( n = 10) or saline ( n = 10) was oral feed three times into CIA in a week. Mice were sacrificed on day 70 after first immunization. (a) Clinical arthritis scores were determined. (b) The joint tissues from CIA: metformin-treated CIA mice were stained with H E, safranin O, and toluidine blue (original magnification, ×200). The average histopathological score is shown in bar graphs (below) (scale bar = 100 μ m). (c) Immunohistochemical detection of IL-17, IL-6, and TNF- α was stained in the synovium of CIA and metformin-treated CIA. All tissues were counterstained with hematoxylin (original magnification, ×400). All images were obtained for each mouse ( n = 10), showing representative images (scale bar = 50 μ m). (d) Mice serum was obtained on day 30 after CII immunization. The serum obtained after first immunization. The levels of <t>IgG,</t> <t>IgG1,</t> and IgG2a antibodies were measured from each group. Mean ± SD of three independent experiments (* P
    Mouse Igg, supplied by Bethyl, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bethyl igg2b
    Flow cytometric analysis of lymphocyte populations in BCAP −/− mice. (A) Expression of BCAP in splenic B cells. Spleen cells stained for CD3 and B220 were analyzed by flow cytometric intracellular staining with an antiserum to BCAP plus goat anti–rabbit <t>IgG-FITC</t> (thick solid line, wild-type mice; shaded area, BCAP −/− mice). Data shown are representative of three independent experiments. (B) Single-cell suspensions from thymus (Thy), bone marrow (BM), lymph node (LN), spleen (SP), and peritoneal cavity (PerC), were stained with the indicated Abs and analyzed using a FACScan ® (wt, wild-type mice). Numbers indicate the percentages of lymphoid cells in the quadrants or enclosed areas. IgM versus IgD and CD21 versus CD23 profiles are shown for B220 + cells, and CD11b versus CD16 profile for B220 − cells. Data shown are representative of six independent experiments.
    Igg2b, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/igg2b/product/Bethyl
    Average 93 stars, based on 1 article reviews
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    Serum IgG1, IgG2, and cytokine expression analysis. ( A ) Anti-IsdA-CTA 2 /B IgG1, ( B ) anti-IsdA-CTA 2 /B IgG2, ( C ) anti-ClfA-CTA 2 /B IgG1, and ( D ) anti-ClfA-CTA 2 /B IgG2 responses in serum on days 14, 20, and 30. Results are reported as ELISA ratios of day X/day 1 at O.D. 370 at serum dilutions of 1:1600. Shown are mean and standard error by treatment with control (gray) and vaccinated (blue) ( n = 3 per group). The log10 of the values were analyzed using analysis of variance (ANOVA), with a compound symmetric covariance structure for cows across days. Model-based estimates were compared between groups within days and adjusted for multipl e comparisons. ( E ) IL-10, IL-6, and IFN-γ expression as determined by quantitative RT-PCR of peripheral blood mononuclear cells (PBMCs) isolated from whole blood after boost on day 20. Results are shown as relative gene expression to GAPDH (2 −ΔΔCt ). Data are presented as mean and standard error of control (gray) and vaccinated (blue) showing median and range ( n = 3 per group). Data were analyzed using a two-group t -test between vaccinated and control. Significant differences between groups are represented as p ≤ 0.05 (*).

    Journal: Vaccines

    Article Title: Evaluation of the Efficacy of a Cholera Toxin-Based Staphylococcus aureus Vaccine against Bovine Intramammary Challenge

    doi: 10.3390/vaccines9010006

    Figure Lengend Snippet: Serum IgG1, IgG2, and cytokine expression analysis. ( A ) Anti-IsdA-CTA 2 /B IgG1, ( B ) anti-IsdA-CTA 2 /B IgG2, ( C ) anti-ClfA-CTA 2 /B IgG1, and ( D ) anti-ClfA-CTA 2 /B IgG2 responses in serum on days 14, 20, and 30. Results are reported as ELISA ratios of day X/day 1 at O.D. 370 at serum dilutions of 1:1600. Shown are mean and standard error by treatment with control (gray) and vaccinated (blue) ( n = 3 per group). The log10 of the values were analyzed using analysis of variance (ANOVA), with a compound symmetric covariance structure for cows across days. Model-based estimates were compared between groups within days and adjusted for multipl e comparisons. ( E ) IL-10, IL-6, and IFN-γ expression as determined by quantitative RT-PCR of peripheral blood mononuclear cells (PBMCs) isolated from whole blood after boost on day 20. Results are shown as relative gene expression to GAPDH (2 −ΔΔCt ). Data are presented as mean and standard error of control (gray) and vaccinated (blue) showing median and range ( n = 3 per group). Data were analyzed using a two-group t -test between vaccinated and control. Significant differences between groups are represented as p ≤ 0.05 (*).

    Article Snippet: After washing, plates were incubated with horseradish peroxidase (HRP)-conjugated anti-bovine IgG, IgG1, IgG2, or IgA (1:10,000 Bethyl Laboratories, Montgomery, TX, USA) at 37 °C for 1 h. Plates were developed with tetramethylbenzidine (PromegaTM TMB One, Thermo Fisher, Waltham, MA, USA) and read at 370 nm per TMB manufacturer’s instruction.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Isolation

    Immunoglobulin G (IgG) antibody responses in serum and milk as determined by enzyme-linked immunosorbent assay (ELISA). Anti-IsdA-CTA 2 /B IgG responses in ( A ) serum and ( B ) milk, anti-ClfA-CTA 2 /B IgG responses in ( C ) serum and ( D ) milk, and anti-CTA 2 /B IgG responses in ( E ) serum and ( F ) milk. Serum was analyzed on days 14, 20, and 30 and milk on days 14, 20, 22, 24, 26, 28, and 30 during the trial period. Results are reported as ELISA ratios of day X/day 1 at O.D. 370 at serum dilutions of 1:1600 and milk dilutions of 1:160. Shown are mean and standard error by treatment with control (gray) and vaccinated (blue) ( n = 3 per group). Significant differences between groups are represented as p ≤ 0.05 (*). The log10 of the values were analyzed using repeated measures analysis of variance (ANOVA) with a compound symmetric covariance structure for cows across days. Model-based estimates were compared between groups within days and adjusted for multiple comparisons.

    Journal: Vaccines

    Article Title: Evaluation of the Efficacy of a Cholera Toxin-Based Staphylococcus aureus Vaccine against Bovine Intramammary Challenge

    doi: 10.3390/vaccines9010006

    Figure Lengend Snippet: Immunoglobulin G (IgG) antibody responses in serum and milk as determined by enzyme-linked immunosorbent assay (ELISA). Anti-IsdA-CTA 2 /B IgG responses in ( A ) serum and ( B ) milk, anti-ClfA-CTA 2 /B IgG responses in ( C ) serum and ( D ) milk, and anti-CTA 2 /B IgG responses in ( E ) serum and ( F ) milk. Serum was analyzed on days 14, 20, and 30 and milk on days 14, 20, 22, 24, 26, 28, and 30 during the trial period. Results are reported as ELISA ratios of day X/day 1 at O.D. 370 at serum dilutions of 1:1600 and milk dilutions of 1:160. Shown are mean and standard error by treatment with control (gray) and vaccinated (blue) ( n = 3 per group). Significant differences between groups are represented as p ≤ 0.05 (*). The log10 of the values were analyzed using repeated measures analysis of variance (ANOVA) with a compound symmetric covariance structure for cows across days. Model-based estimates were compared between groups within days and adjusted for multiple comparisons.

    Article Snippet: After washing, plates were incubated with horseradish peroxidase (HRP)-conjugated anti-bovine IgG, IgG1, IgG2, or IgA (1:10,000 Bethyl Laboratories, Montgomery, TX, USA) at 37 °C for 1 h. Plates were developed with tetramethylbenzidine (PromegaTM TMB One, Thermo Fisher, Waltham, MA, USA) and read at 370 nm per TMB manufacturer’s instruction.

    Techniques: Enzyme-linked Immunosorbent Assay

    Rapamycin suppresses T cell alloreactivity, proinflammatory cytokines, and production of immunoglobulin and induces mitochondrial dysfunction in vitro. a In a mixed lymphocyte reaction assay, B/c splenic T cells (responders) were incubated with irradiated B/c splenic antigen-presenting cells for 4 days. Responder cells were cultured in the presence or absence of rapamycin. b IFN-γ and IL-17 levels in the supernatants, as measured by enzyme-linked immunosorbent assay (ELISA). c IL-17, IL-6, TNF-α, and IgG concentrations in culture supernatants in the presence of anti-CD3 (0.5 μg/ml) or lipopolysaccharide (100 ng/ml) with rapamycin 100 nM, as determined by ELISA. d The Treg population, as determined by flow cytometry. e JC-1 populations in isolated splenic cells, as determined by flow cytometry. The data are representative of at least three independent experiments. Data represent the mean ± SD of three independent experiments. ** P

    Journal: Arthritis Research & Therapy

    Article Title: Metformin rescues rapamycin-induced mitochondrial dysfunction and attenuates rheumatoid arthritis with metabolic syndrome

    doi: 10.1186/s13075-020-02174-3

    Figure Lengend Snippet: Rapamycin suppresses T cell alloreactivity, proinflammatory cytokines, and production of immunoglobulin and induces mitochondrial dysfunction in vitro. a In a mixed lymphocyte reaction assay, B/c splenic T cells (responders) were incubated with irradiated B/c splenic antigen-presenting cells for 4 days. Responder cells were cultured in the presence or absence of rapamycin. b IFN-γ and IL-17 levels in the supernatants, as measured by enzyme-linked immunosorbent assay (ELISA). c IL-17, IL-6, TNF-α, and IgG concentrations in culture supernatants in the presence of anti-CD3 (0.5 μg/ml) or lipopolysaccharide (100 ng/ml) with rapamycin 100 nM, as determined by ELISA. d The Treg population, as determined by flow cytometry. e JC-1 populations in isolated splenic cells, as determined by flow cytometry. The data are representative of at least three independent experiments. Data represent the mean ± SD of three independent experiments. ** P

    Article Snippet: Measurement of immunoglobulin G concentrations The immunoglobulin G (IgG) concentrations were measured using mouse IgG ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX, USA) in splenocyte culture supernatant.

    Techniques: In Vitro, Incubation, Irradiation, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Isolation

    Therapeutic effects of metformin in CIA model. CIA was induced in C57BL/6 mice. Metformin 5 mg/mice ( n = 10) or saline ( n = 10) was oral feed three times into CIA in a week. Mice were sacrificed on day 70 after first immunization. (a) Clinical arthritis scores were determined. (b) The joint tissues from CIA: metformin-treated CIA mice were stained with H E, safranin O, and toluidine blue (original magnification, ×200). The average histopathological score is shown in bar graphs (below) (scale bar = 100 μ m). (c) Immunohistochemical detection of IL-17, IL-6, and TNF- α was stained in the synovium of CIA and metformin-treated CIA. All tissues were counterstained with hematoxylin (original magnification, ×400). All images were obtained for each mouse ( n = 10), showing representative images (scale bar = 50 μ m). (d) Mice serum was obtained on day 30 after CII immunization. The serum obtained after first immunization. The levels of IgG, IgG1, and IgG2a antibodies were measured from each group. Mean ± SD of three independent experiments (* P

    Journal: Mediators of Inflammation

    Article Title: Metformin Attenuates Experimental Autoimmune Arthritis through Reciprocal Regulation of Th17/Treg Balance and Osteoclastogenesis

    doi: 10.1155/2014/973986

    Figure Lengend Snippet: Therapeutic effects of metformin in CIA model. CIA was induced in C57BL/6 mice. Metformin 5 mg/mice ( n = 10) or saline ( n = 10) was oral feed three times into CIA in a week. Mice were sacrificed on day 70 after first immunization. (a) Clinical arthritis scores were determined. (b) The joint tissues from CIA: metformin-treated CIA mice were stained with H E, safranin O, and toluidine blue (original magnification, ×200). The average histopathological score is shown in bar graphs (below) (scale bar = 100 μ m). (c) Immunohistochemical detection of IL-17, IL-6, and TNF- α was stained in the synovium of CIA and metformin-treated CIA. All tissues were counterstained with hematoxylin (original magnification, ×400). All images were obtained for each mouse ( n = 10), showing representative images (scale bar = 50 μ m). (d) Mice serum was obtained on day 30 after CII immunization. The serum obtained after first immunization. The levels of IgG, IgG1, and IgG2a antibodies were measured from each group. Mean ± SD of three independent experiments (* P

    Article Snippet: Measurement of Immunoglobulin (Ig) Concentrations Anti-IgG, IgG1, and IgG2a were measured by mouse IgG, IgG1, and IgG2a ELISA quantitation kits (Bethyl Laboratories, Montgomery, TX) [ ].

    Techniques: Mouse Assay, Staining, Immunohistochemistry

    Flow cytometric analysis of lymphocyte populations in BCAP −/− mice. (A) Expression of BCAP in splenic B cells. Spleen cells stained for CD3 and B220 were analyzed by flow cytometric intracellular staining with an antiserum to BCAP plus goat anti–rabbit IgG-FITC (thick solid line, wild-type mice; shaded area, BCAP −/− mice). Data shown are representative of three independent experiments. (B) Single-cell suspensions from thymus (Thy), bone marrow (BM), lymph node (LN), spleen (SP), and peritoneal cavity (PerC), were stained with the indicated Abs and analyzed using a FACScan ® (wt, wild-type mice). Numbers indicate the percentages of lymphoid cells in the quadrants or enclosed areas. IgM versus IgD and CD21 versus CD23 profiles are shown for B220 + cells, and CD11b versus CD16 profile for B220 − cells. Data shown are representative of six independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: Essential Immunoregulatory Role for BCAP in B Cell Development and Function

    doi: 10.1084/jem.20011751

    Figure Lengend Snippet: Flow cytometric analysis of lymphocyte populations in BCAP −/− mice. (A) Expression of BCAP in splenic B cells. Spleen cells stained for CD3 and B220 were analyzed by flow cytometric intracellular staining with an antiserum to BCAP plus goat anti–rabbit IgG-FITC (thick solid line, wild-type mice; shaded area, BCAP −/− mice). Data shown are representative of three independent experiments. (B) Single-cell suspensions from thymus (Thy), bone marrow (BM), lymph node (LN), spleen (SP), and peritoneal cavity (PerC), were stained with the indicated Abs and analyzed using a FACScan ® (wt, wild-type mice). Numbers indicate the percentages of lymphoid cells in the quadrants or enclosed areas. IgM versus IgD and CD21 versus CD23 profiles are shown for B220 + cells, and CD11b versus CD16 profile for B220 − cells. Data shown are representative of six independent experiments.

    Article Snippet: Total serum levels of IgM, IgG1, IgG2a, IgG2b, IgG3, and IgA were determined using ELISA quantitation kits (Bethyl Lab.).

    Techniques: Flow Cytometry, Mouse Assay, Expressing, Staining