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  • 99
    Thermo Fisher anti cd28
    MicroRNA‐126 (Mir‐126) deficiency altered the expression of cytokines in CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) were cultured in the presence of anti‐CD3 (20 <t>μg/ml)/anti‐CD28</t> (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml); 72 h later, the relative expression of cytokines including IL‐4, IL‐6, IL‐10, IL‐12, transforming growth factor (TGF)‐β, tumour necrosis factor (TNF)‐α and interferon (IFN)‐γ were detected by real‐time polymerase chain reaction (RT–PCR) and calculated (a). (b) The expression levels of IFN‐γ, IL‐4 and IL‐17A in CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) and calculated (c). One representative example of three independent experiments is shown. * P
    Anti Cd28, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti cd28
    MicroRNA‐126 (Mir‐126) deficiency altered the expression of cytokines in CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) were cultured in the presence of anti‐CD3 (20 <t>μg/ml)/anti‐CD28</t> (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml); 72 h later, the relative expression of cytokines including IL‐4, IL‐6, IL‐10, IL‐12, transforming growth factor (TGF)‐β, tumour necrosis factor (TNF)‐α and interferon (IFN)‐γ were detected by real‐time polymerase chain reaction (RT–PCR) and calculated (a). (b) The expression levels of IFN‐γ, IL‐4 and IL‐17A in CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) and calculated (c). One representative example of three independent experiments is shown. * P
    Anti Cd28, supplied by BioLegend, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd28/product/BioLegend
    Average 97 stars, based on 1 article reviews
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    Thermo Fisher dynabeads human t activator cd3 cd28
    Changes in chromatin state in <t>human</t> T cell activation ( a ) Experimental overview (left) and schematic of nomenclature (right). ( b ) Differential chromatin accessibility. Regions of open chromatin (columns) in six samples (rows) before (top, Th-specific) and 48hr after (bottom, Th stim- specific) activation of primary T cells with <t>anti-CD3/CD28</t> antibodies. ( c ) Overlap with previously annotated T cell enhancers. For each annotation, expected (x-axis) vs. observed (y-axis) percentages of annotated features overlapping Th-specific (left), Th stim -specific (center) and shared peaks (right). ( d ) Overlap with GWAS variants. For each phenotype or disease, expected (x-axis) vs. observed (y-axis) percentages of GWAS loci overlapping Th-specific (left), Th stim -specific (center), or shared (right) peaks.
    Dynabeads Human T Activator Cd3 Cd28, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dynabeads human t activator cd3 cd28/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    99
    Thermo Fisher anti cd28 antibodies
    SIRT5 deficiency does not affect proliferation of and IL-2 production by splenocytes. SIRT5 +/+ and SIRT5 −/− splenocytes were incubated for 48 h with LPS (5 μg/ml), CpG (2 μg/ml), Pam 3 CSK 4 (5 μg/ml), TSST-1 (2 μg/ml), <t>anti-CD3/CD28</t> antibodies (1μg/ml) and PMA + ionomycin (PMA/iono, 10 ng/ml/100 ng/ml). (A) Proliferation was measured by 3 H-thymidine incorporation. (B) IL-2 concentrations in cell culture supernatants were quantified by ELISA. Data are means ± SD of one experiment performed with three mice and are representative of two experiments. P > 0.05 for all conditions.
    Anti Cd28 Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd28 antibodies/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    The CD28 2 monoclonal antibody specifically binds with the human 44 kDa homodimeric trans membrane glycoprotein CD28 expressed on the surface of most mature T lymphocytes plasma cells and thymocytes
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    The CD28 Antibody CD28 2 from Novus Biologicals is a mouse monoclonal antibody to CD28 This antibody reacts with human primate The CD28 Antibody CD28 2 has been validated for
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    CD28 molecule Recombinant Protein Epitope Signature Tag PrEST antigen sequence
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    CD28 Antibody is a primary antibody against CD28 Please contact us at lt a href mailto info abbexa com gt info abbexa com lt a gt for stock information before
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    Image Search Results


    MicroRNA‐126 (Mir‐126) deficiency altered the expression of cytokines in CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml); 72 h later, the relative expression of cytokines including IL‐4, IL‐6, IL‐10, IL‐12, transforming growth factor (TGF)‐β, tumour necrosis factor (TNF)‐α and interferon (IFN)‐γ were detected by real‐time polymerase chain reaction (RT–PCR) and calculated (a). (b) The expression levels of IFN‐γ, IL‐4 and IL‐17A in CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) and calculated (c). One representative example of three independent experiments is shown. * P

    Journal: Clinical and Experimental Immunology

    Article Title: MicroRNA‐126 deficiency enhanced the activation and function of CD4+T cells by elevating IRS‐1 pathway

    doi: 10.1111/cei.13067

    Figure Lengend Snippet: MicroRNA‐126 (Mir‐126) deficiency altered the expression of cytokines in CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml); 72 h later, the relative expression of cytokines including IL‐4, IL‐6, IL‐10, IL‐12, transforming growth factor (TGF)‐β, tumour necrosis factor (TNF)‐α and interferon (IFN)‐γ were detected by real‐time polymerase chain reaction (RT–PCR) and calculated (a). (b) The expression levels of IFN‐γ, IL‐4 and IL‐17A in CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) and calculated (c). One representative example of three independent experiments is shown. * P

    Article Snippet: Coating with 20 μg/ml anti‐CD3e (eBioscience, San Diego, CA, USA; 16‐0031‐86) for 4°C overnight in plates and cells were co‐stimulated with 4 ng/ml anti‐CD28 (eBioscience; 16‐0281‐85) antibody and 10 ng/ml plus interleukin (IL)‐2 (ProSpec, cyt‐370‐b).

    Techniques: Expressing, Purification, FACS, Magnetic Cell Separation, Mouse Assay, Cell Culture, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Fluorescence

    MicroRNA‐126 (Mir‐126) deficiency enhanced the activation and proliferation of CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) respectively. Next, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 72 h. Then, the relative expression of miR‐126 was detected by real‐time polymerase chain reaction (RT–PCR) assay (a). The expression levels of CD69, CD62L and CD44 on CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) (b) and calculated (c). Proliferation of CD4 + T cells from miR‐126KD mice and WT mice were assessed by Ki‐67 staining (d) and calculated (e). (f) The apoptosis of CD4 + T cells was also analysed by FACS and calculated (g). One representative example of three independent experiments is shown. * P

    Journal: Clinical and Experimental Immunology

    Article Title: MicroRNA‐126 deficiency enhanced the activation and function of CD4+T cells by elevating IRS‐1 pathway

    doi: 10.1111/cei.13067

    Figure Lengend Snippet: MicroRNA‐126 (Mir‐126) deficiency enhanced the activation and proliferation of CD4 + T cells. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from splenocytes in miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6) respectively. Next, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 72 h. Then, the relative expression of miR‐126 was detected by real‐time polymerase chain reaction (RT–PCR) assay (a). The expression levels of CD69, CD62L and CD44 on CD4 + T cells were analysed by fluorescence activated cell sorting (FACS) (b) and calculated (c). Proliferation of CD4 + T cells from miR‐126KD mice and WT mice were assessed by Ki‐67 staining (d) and calculated (e). (f) The apoptosis of CD4 + T cells was also analysed by FACS and calculated (g). One representative example of three independent experiments is shown. * P

    Article Snippet: Coating with 20 μg/ml anti‐CD3e (eBioscience, San Diego, CA, USA; 16‐0031‐86) for 4°C overnight in plates and cells were co‐stimulated with 4 ng/ml anti‐CD28 (eBioscience; 16‐0281‐85) antibody and 10 ng/ml plus interleukin (IL)‐2 (ProSpec, cyt‐370‐b).

    Techniques: Activation Assay, Purification, FACS, Magnetic Cell Separation, Mouse Assay, Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Staining

    The effect of microRNA‐126 (Mir‐126) deficiency on the transduction of related signalling pathway. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6), respectively, Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. (a) The expression levels of phospho‐extracellular regulated kinase (p‐ERK), phospho‐protein kinase B (p‐AKT) and phospho‐nuclear factor kinase kappa B (p‐NF‐κB) were determined by fluorescence activated cell sorting (FACS) and calculated (b). (c) Schematic representation of the underlying mechanism of miR‐126 deficiency on the activation and function of CD4 + ]

    Journal: Clinical and Experimental Immunology

    Article Title: MicroRNA‐126 deficiency enhanced the activation and function of CD4+T cells by elevating IRS‐1 pathway

    doi: 10.1111/cei.13067

    Figure Lengend Snippet: The effect of microRNA‐126 (Mir‐126) deficiency on the transduction of related signalling pathway. CD4 + CD62L + T cells purified by magnetic‐activated cell sorting (MACS) from miR‐126 knock‐down (KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6), respectively, Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. (a) The expression levels of phospho‐extracellular regulated kinase (p‐ERK), phospho‐protein kinase B (p‐AKT) and phospho‐nuclear factor kinase kappa B (p‐NF‐κB) were determined by fluorescence activated cell sorting (FACS) and calculated (b). (c) Schematic representation of the underlying mechanism of miR‐126 deficiency on the activation and function of CD4 + ]

    Article Snippet: Coating with 20 μg/ml anti‐CD3e (eBioscience, San Diego, CA, USA; 16‐0031‐86) for 4°C overnight in plates and cells were co‐stimulated with 4 ng/ml anti‐CD28 (eBioscience; 16‐0281‐85) antibody and 10 ng/ml plus interleukin (IL)‐2 (ProSpec, cyt‐370‐b).

    Techniques: Transduction, Purification, FACS, Magnetic Cell Separation, Mouse Assay, Cell Culture, Expressing, Fluorescence, Activation Assay

    The expression of insulin receptor substrate 1 (IRS‐1) in CD4 + T cells. CD4 + CD62L + T cells were purified by magnetic‐activated cell sorting (MACS) from microRNA‐126 knock‐down (Mir‐126KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6), respectively. Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. The relative expression of indicated genes, including phosphomannomutase 1 (PMM1), target of Myb1 membrane trafficking protein (Tom1), phosphatidylinositol‐4,5‐bisphosphate 3‐kinase catalytic subunit delta (PIK3CD), regulator of G protein signalling 3 (RGS3), tuberous sclerosis 1 (TSC1), insulin receptor substrate 1 (IRS‐1), ADAM metallopeptidase domain 9 (ADAM9), EGF‐like domain multiple 7 (EGFL7), vascular cell adhesion molecule 1 (VCAM1), PHD finger protein 7 (Phf7), solute carrier family 39 member 6 (SLC39a6), Huntingtin interacting protein 1 (HIP1) and poly(ADP‐ribose) polymerase family member 16 (PARP16P), were analysed by real‐time polymerase chain reaction (RT–PCR) assay and calculated (a). (b) Putative miR‐126‐binding sites in the 3' untranslated region (UTR) of murine IRS‐1. (c) CD4 + CD62L + T cells were purified by MACS from FVB/N6 miR‐126KD mice and WT mice, respectively. Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. The expression of IRS‐1 protein was determined by Western blot and calculated. CD4 + CD62L + T cells were purified by MACS from miR‐126KD mice, Then, cells were transfected with p‐IRS‐1 RNAi (5 ng) and cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus IL‐2 (10 ng/ml) for 48 h. (d) The expression of IRS‐1 was analysed by real‐time polymerase chain reaction (RT–PCR). (e) Call counting kit‐8 assay. The expression levels of CD69 and CD44 were analysed by fluorescence activated cell sorting (FACS) and the percentage was calculated (f–i). (j) The expression levels of interferon (IFN)‐γ and IL‐4 were analysed by FACS and the percentage was calculated (k). One representative example of three independent experiments is shown. * P

    Journal: Clinical and Experimental Immunology

    Article Title: MicroRNA‐126 deficiency enhanced the activation and function of CD4+T cells by elevating IRS‐1 pathway

    doi: 10.1111/cei.13067

    Figure Lengend Snippet: The expression of insulin receptor substrate 1 (IRS‐1) in CD4 + T cells. CD4 + CD62L + T cells were purified by magnetic‐activated cell sorting (MACS) from microRNA‐126 knock‐down (Mir‐126KD) mice and wild‐type (WT) mice (8–10 weeks old, n = 6), respectively. Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. The relative expression of indicated genes, including phosphomannomutase 1 (PMM1), target of Myb1 membrane trafficking protein (Tom1), phosphatidylinositol‐4,5‐bisphosphate 3‐kinase catalytic subunit delta (PIK3CD), regulator of G protein signalling 3 (RGS3), tuberous sclerosis 1 (TSC1), insulin receptor substrate 1 (IRS‐1), ADAM metallopeptidase domain 9 (ADAM9), EGF‐like domain multiple 7 (EGFL7), vascular cell adhesion molecule 1 (VCAM1), PHD finger protein 7 (Phf7), solute carrier family 39 member 6 (SLC39a6), Huntingtin interacting protein 1 (HIP1) and poly(ADP‐ribose) polymerase family member 16 (PARP16P), were analysed by real‐time polymerase chain reaction (RT–PCR) assay and calculated (a). (b) Putative miR‐126‐binding sites in the 3' untranslated region (UTR) of murine IRS‐1. (c) CD4 + CD62L + T cells were purified by MACS from FVB/N6 miR‐126KD mice and WT mice, respectively. Then, cells were cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus interleukin (IL)‐2 (10 ng/ml) for 48 h. The expression of IRS‐1 protein was determined by Western blot and calculated. CD4 + CD62L + T cells were purified by MACS from miR‐126KD mice, Then, cells were transfected with p‐IRS‐1 RNAi (5 ng) and cultured in the presence of anti‐CD3 (20 μg/ml)/anti‐CD28 (4 ng/ml) antibody plus IL‐2 (10 ng/ml) for 48 h. (d) The expression of IRS‐1 was analysed by real‐time polymerase chain reaction (RT–PCR). (e) Call counting kit‐8 assay. The expression levels of CD69 and CD44 were analysed by fluorescence activated cell sorting (FACS) and the percentage was calculated (f–i). (j) The expression levels of interferon (IFN)‐γ and IL‐4 were analysed by FACS and the percentage was calculated (k). One representative example of three independent experiments is shown. * P

    Article Snippet: Coating with 20 μg/ml anti‐CD3e (eBioscience, San Diego, CA, USA; 16‐0031‐86) for 4°C overnight in plates and cells were co‐stimulated with 4 ng/ml anti‐CD28 (eBioscience; 16‐0281‐85) antibody and 10 ng/ml plus interleukin (IL)‐2 (ProSpec, cyt‐370‐b).

    Techniques: Expressing, Purification, FACS, Magnetic Cell Separation, Mouse Assay, Cell Culture, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Western Blot, Transfection, Fluorescence

    Changes in chromatin state in human T cell activation ( a ) Experimental overview (left) and schematic of nomenclature (right). ( b ) Differential chromatin accessibility. Regions of open chromatin (columns) in six samples (rows) before (top, Th-specific) and 48hr after (bottom, Th stim- specific) activation of primary T cells with anti-CD3/CD28 antibodies. ( c ) Overlap with previously annotated T cell enhancers. For each annotation, expected (x-axis) vs. observed (y-axis) percentages of annotated features overlapping Th-specific (left), Th stim -specific (center) and shared peaks (right). ( d ) Overlap with GWAS variants. For each phenotype or disease, expected (x-axis) vs. observed (y-axis) percentages of GWAS loci overlapping Th-specific (left), Th stim -specific (center), or shared (right) peaks.

    Journal: Nature genetics

    Article Title: Genetic determinants of co-accessible chromatin regions in activated T cells across humans

    doi: 10.1038/s41588-018-0156-2

    Figure Lengend Snippet: Changes in chromatin state in human T cell activation ( a ) Experimental overview (left) and schematic of nomenclature (right). ( b ) Differential chromatin accessibility. Regions of open chromatin (columns) in six samples (rows) before (top, Th-specific) and 48hr after (bottom, Th stim- specific) activation of primary T cells with anti-CD3/CD28 antibodies. ( c ) Overlap with previously annotated T cell enhancers. For each annotation, expected (x-axis) vs. observed (y-axis) percentages of annotated features overlapping Th-specific (left), Th stim -specific (center) and shared peaks (right). ( d ) Overlap with GWAS variants. For each phenotype or disease, expected (x-axis) vs. observed (y-axis) percentages of GWAS loci overlapping Th-specific (left), Th stim -specific (center), or shared (right) peaks.

    Article Snippet: Cells were either left untreated or stimulated with beads conjugated with anti-CD3 and anti-CD28 antibodies (Dynabeads, Invitrogen #11131D, Life Technologies) at a cell:bead ratio of 1:1 for 48 hours, a time point we previously found to maximize the gene expression response in CD4+ T cells.

    Techniques: Activation Assay, GWAS

    SIRT5 deficiency does not affect proliferation of and IL-2 production by splenocytes. SIRT5 +/+ and SIRT5 −/− splenocytes were incubated for 48 h with LPS (5 μg/ml), CpG (2 μg/ml), Pam 3 CSK 4 (5 μg/ml), TSST-1 (2 μg/ml), anti-CD3/CD28 antibodies (1μg/ml) and PMA + ionomycin (PMA/iono, 10 ng/ml/100 ng/ml). (A) Proliferation was measured by 3 H-thymidine incorporation. (B) IL-2 concentrations in cell culture supernatants were quantified by ELISA. Data are means ± SD of one experiment performed with three mice and are representative of two experiments. P > 0.05 for all conditions.

    Journal: Frontiers in Immunology

    Article Title: Sirtuin 5 Deficiency Does Not Compromise Innate Immune Responses to Bacterial Infections

    doi: 10.3389/fimmu.2018.02675

    Figure Lengend Snippet: SIRT5 deficiency does not affect proliferation of and IL-2 production by splenocytes. SIRT5 +/+ and SIRT5 −/− splenocytes were incubated for 48 h with LPS (5 μg/ml), CpG (2 μg/ml), Pam 3 CSK 4 (5 μg/ml), TSST-1 (2 μg/ml), anti-CD3/CD28 antibodies (1μg/ml) and PMA + ionomycin (PMA/iono, 10 ng/ml/100 ng/ml). (A) Proliferation was measured by 3 H-thymidine incorporation. (B) IL-2 concentrations in cell culture supernatants were quantified by ELISA. Data are means ± SD of one experiment performed with three mice and are representative of two experiments. P > 0.05 for all conditions.

    Article Snippet: Stimuli were Salmonella minnesota ultra pure LPS (InvivoGen, San Diego, CA), Pam3 CSK4 (EMC microcollections, Tübingen, Germany), CpG ODN 1826 (CpG, InvivoGen), toxic shock syndrome toxin-1 (TSST-1, Toxin Technology, Sarasota, FL), concanavalin A (Sigma-Aldrich, St. Louis, MI), anti-CD3ε, and anti-CD28 antibodies (clones 145-2C11 and 37.51, eBioscience, San Diego, CA) and phorbol-12-myristate-13-acetate (PMA) plus ionomycin (Sigma-Aldrich) or bacteria.

    Techniques: Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Mouse Assay