polyclonal goat anti human il 1ra antibody  (R&D Systems)

Journal: Journal of leukocyte biology

Article Title: Interleukin-15 delays human neutrophil apoptosis by intracellular events and not via extracellular factors: role of Mcl-1 and decreased activity of caspase-3 and caspase-8.

doi: 10.1189/jlb.1103585

Figure Lengend Snippet: Fig. 1. IL-15 induces de novo protein synthe- sis of a neutrophil protein identified as IL-1Ra. Human neutrophils were metabolically labeled (A and C) or not (panel B) as described in Materials and Methods and incubated for 24 h in the presence or absence of stimuli. Extracel- lular fractions (supernatants) were prepared as described in Materials and Methods and then run on 15% SDS-PAGE. Ctrl, Unstimulated cells; GM, 65 ng/ml GM-CSF; CHX, 10 g/ml CHX; and IL-15, 250 ng/ml IL-15. Note the very weak signal obtained after CHX treatment on the film (35S; A) when compared with the total protein loaded in the corresponding lane (Coomassie blue). In other experiments, super- natants were harvested, prepared, and immuno- blotting (B), or immunoprecipitation (IP; C) was performed with an anti-human IL-1Ra antibody as described in Materials and Methods. Results are from one representative experiment of at least three. mw, Molecular weight.

Article Snippet: Recombinant human (rh)IL-1Ra and the goat anti-human IL-1Ra polyclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Metabolic Labelling, Labeling, Incubation, SDS Page, Immunoprecipitation, Molecular Weight

Journal: Journal of leukocyte biology

Article Title: Interleukin-15 delays human neutrophil apoptosis by intracellular events and not via extracellular factors: role of Mcl-1 and decreased activity of caspase-3 and caspase-8.

doi: 10.1189/jlb.1103585

Figure Lengend Snippet: Fig. 2. Quantification of IL-1Ra in the intracellular and extracellular fractions from IL-15-induced neutrophils. Neutrophils were incubated for 24 h in the presence of buffer (C), LPS, GM-CSF (GM), or IL-15 (100 or 250 ng/ml), and intra- and extracellular fractions were pre- pared for quantification of IL-1Ra, IL-1, and IL-1 by ELISA, as described in Materials and Methods. Results are mean SEM (from eight different blood donors). *, P 0.05, by ANOVA.

Article Snippet: Recombinant human (rh)IL-1Ra and the goat anti-human IL-1Ra polyclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Journal of leukocyte biology

Article Title: Interleukin-15 delays human neutrophil apoptosis by intracellular events and not via extracellular factors: role of Mcl-1 and decreased activity of caspase-3 and caspase-8.

doi: 10.1189/jlb.1103585

Figure Lengend Snippet: Fig. 3. IL-1Ra does not modulate the apoptotic rate of human neutrophils. Freshly isolated human neutrophils (10106 cells/ml in RPMI 1640 supple- mented with 10% FCS) were incubated for 24 h in the presence of buffer (Ctrl), GM-CSF (GM), IL-15, or increasing concentrations of IL-1Ra (10, 100, or 500 ng/ml), and apoptosis was evaluated by flow cytometry (FITC–Annexin-V and FITC–CD16) and by cytology as described in Materials and Methods. Results are mean SEM (n3). *, P 0.05, by ANOVA.

Article Snippet: Recombinant human (rh)IL-1Ra and the goat anti-human IL-1Ra polyclonal antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Isolation, Incubation, Cytometry

Journal: Human gene therapy

Article Title: Cytokine gene therapy for myocarditis by in vivo electroporation.

doi: 10.1089/104303401750270940

Figure Lengend Snippet: FIG. 1. Immunoblot of the culture supernatant from BMT-10 cells transfected with pCAGGS-IL-1ra. Goat anti-IL-1ra poly- clonal Ab detected a protein of 17 kDa, which is the expected size for mouse IL-1ra. The lanes represent supernatants from pCAGGS-transfected and pCAGGS-IL-1ra-transfected cells, and recombinant murine IL-1ra.

Article Snippet: The membrane was incubated at room temperature for 3 hr with goat anti-IL-1ra polyclonal Ab (R&D Systems, Minneapolis, MN), and washed.

Techniques: Western Blot, Transfection, Recombinant

Journal: Human gene therapy

Article Title: Cytokine gene therapy for myocarditis by in vivo electroporation.

doi: 10.1089/104303401750270940

Figure Lengend Snippet: FIG. 2. (A) Time course of the serum concentration of IL-1ra after pCAGGS-IL-1ra transfer to uninfected mice by in vivo elec- troporation. The serum IL-1ra concentration peaked at day 5 (solid squares). Transfer of the empty pCAGGS vector did not sig- nificantly affect the serum IL-1ra levels (solid diamonds). (B) Time course of the serum concentration of IL-1ra after EMCV in- oculation and pCAGGS-IL-1ra transfer. The serum IL-1ra levels were elevated by EMCV inoculation (solid circles) and were further increased by pCAGGS-IL-1ra transfer by in vivo electroporation (open squares) (n 5 3, each). Intramuscular injection of pCAGGS-IL-1ra without electroporation (open circles) or the empty vector with electroporation (open triangles) showed only marginal effects on the IL-1ra levels in EMCV-inoculated mice.

Article Snippet: The membrane was incubated at room temperature for 3 hr with goat anti-IL-1ra polyclonal Ab (R&D Systems, Minneapolis, MN), and washed.

Techniques: Concentration Assay, In Vivo, Plasmid Preparation, Electroporation, Injection

Journal: Human gene therapy

Article Title: Cytokine gene therapy for myocarditis by in vivo electroporation.

doi: 10.1089/104303401750270940

Figure Lengend Snippet: FIG. 3. Survival curve after EMCV inoculation. Analysis of the survival curve revealed that the pCAGGS-IL-1ra-treated group (n 5 15) showed significantly higher survival than the empty vector-treated group (n 5 15) and PBS-treated group (n 5 10) (p , 0.05). vIL-10 gene transfer (n 5 15) also decreased the mortality of mice inoculated with EMCV (p , 0.005).

Article Snippet: The membrane was incubated at room temperature for 3 hr with goat anti-IL-1ra polyclonal Ab (R&D Systems, Minneapolis, MN), and washed.

Techniques: Plasmid Preparation

Journal: Human gene therapy

Article Title: Cytokine gene therapy for myocarditis by in vivo electroporation.

doi: 10.1089/104303401750270940

Figure Lengend Snippet: FIG. 4. Histological examination. The quantitative pathological scores of myocardial infiltration revealed significant improve- ment in both the IL-1ra-treated and vIL-10-treated groups (n 5 5, each) (IL-1ra-treated group vs. empty vector-treated group, p , 0.01; vIL-10-treated group vs. empty vector-treated group, p , 0.01). Bottom: Representative histological results for each group.

Article Snippet: The membrane was incubated at room temperature for 3 hr with goat anti-IL-1ra polyclonal Ab (R&D Systems, Minneapolis, MN), and washed.

Techniques: Plasmid Preparation

Journal: Human gene therapy

Article Title: Cytokine gene therapy for myocarditis by in vivo electroporation.

doi: 10.1089/104303401750270940

Figure Lengend Snippet: FIG. 5. Cytokine expression in the heart. Total RNA was extracted from the hearts of mice of the IL-1ra-, pCAGGS-IL-10, and empty vector-treated groups. RNA was reverse transcribed and subjected to real-time quantitative PCR to determine the lev- els of mRNA of cytokines in the heart. (A) In the pCAGGS-IL-1ra-treated mice, the expression levels of TNF-a and iNOS in the heart were significantly decreased (n 5 5, each). (B) In pCAGGS-vIL-10-treated mice, the expression levels of IFN-g and iNOS in the heart were significantly decreased (n 5 5, each).

Article Snippet: The membrane was incubated at room temperature for 3 hr with goat anti-IL-1ra polyclonal Ab (R&D Systems, Minneapolis, MN), and washed.

Techniques: Expressing, Plasmid Preparation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: The Journal of Neuroscience

Article Title: Inflammatory Genes Are Upregulated in Expanded Ataxin-3-Expressing Cell Lines and Spinocerebellar Ataxia Type 3 Brains

doi: 10.1523/JNEUROSCI.21-15-05389.2001

Figure Lengend Snippet: Altered expression of the inflammatory mediators IL-1ra, IL-1β, and SDF1 and increased numbers of CD68- and IL-1β-immunoreactive glial cells in pons sections of diseased SCA3 brain. Comparison of IL-1ra-immunostained control (A) and disease pons sections (B, C) revealed a significantly increased cytoplasmic staining of pontine neurons (B and inset) and several immunoreactive plaque-like structures (C) in SCA3 patients. Comparison of IL-1β-immunostained control (D) and disease pons sections (E) showed an enhanced staining of pontine neurons (E andinset) in SCA3 patients. Immunohistochemical analysis using anti-SDF1 antibodies showed an intense staining of pontine neurons in SCA3 cases (G) that was also present in controls but to a much lesser extent (F). Immunostaining for CD68 revealed several positive perineuronal cells displaying a typical morphology of activated microglia in SCA3 pons (I), whereas CD68-positive cells were less frequently in controls (H). Immunohistochemical analysis against IL-1β showed a significant increase of reactive astrocytes in SCA3 pons (K) compared with control pons section (J). Tissue sections were counterstained with hematoxylin. Scale bars: A–K, 100 μm; insets, 10 μm.

Article Snippet: The antibodies and dilutions used in this study were as follows: rabbit polyclonal anti-human MMP-2 antibody (1:800; Chemicon, Temecula, CA); mouse monoclonal anti-human Aβ (amino acid residues 1–16) antibody (1:100; Chemicon); goat polyclonal anti-human IL-1ra antibody (1:20; R & D Systems, Minneapolis, MN); goat polyclonal anti-human IL-1β antibody (1:20; R & D Systems); goat polyclonal anti-human SDF1 antibody (1:100; R & D Systems); mouse monoclonal anti-human CD68 antibody (1:25; Dako, Hamburg, Germany); and rabbit polyclonal anti-human glial fibrillary acidic protein (GFAP) (1:800; Dako).

Techniques: Expressing, Comparison, Control, Staining, Immunohistochemical staining, Immunostaining