polyclonal rabbit anti human a2bar antibody  (Alomone Labs)

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: (A-D): Pulmonary epithelial cells (A549) or vascular endothelia (HMEC-1) were exposed to indicated concentrations of inflammatory mediators, and A2BAR transcript levels were determined by real-time RT-PCR following 0–24h of exposure time. Data were calculated relative to the internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (0h of exposure) ± SD at each indicated time (n=4). (E) Degradation curve of A2BAR mRNA in HMEC-1 cells treated with PGE2 and challenged with DRB (n=3; *p<0.005 by ANOVA). (F). A2BAR expression in primary pulmonary endothelial cells treated with IL-6 (HMVEC-L) (n=4). (G) Western-blot analysis of A2BAR protein following 24h of IL-6 stimulation at indicated concentrations. The same blot was stripped and re-probed for murine β-actin to control for loading conditions. One representative of three Western blots is displayed.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Quantitative RT-PCR, Control, Expressing, Western Blot

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: Mice were exposed to 30 min of LPS inhalation, and animals were sacrificed after 4h. (A) Pulmonary A2BAR transcript levels were assessed by real-time RT-PCR. Data were calculated relative to internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (-LPS) ± SD (n=9). Western-blot analysis of A2BAR protein following in vivo exposure to inhaled LPS. The same blot was stripped and re-probed for murine β-actin to control for loading conditions. One representative of four Western blots is displayed. (C) Pulmonary immunohistochemistry for the A2BAR following LPS exposure. Lungs from mice exposed to 30 min of LPS inhalation or vehicle control (Vehicle) were harvested. Sections were stained with antibodies specifical for murine A2BAR (green), or isotype controls. 4′,6-diamidino-2-phenylindole was used for nuclear counterstain (blue) (magnification, 400×). One representative image from 3 pulmonic sections is displayed.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Quantitative RT-PCR, Control, Western Blot, In Vivo, Immunohistochemistry, Staining

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: 8–12 week old C57BL/6J mice matched in age, gender and weight were pre-exposed to 30 min of inhaled A2BAR antagonist (MRS1754) or vehicle, followed by 30 min of LPS or vehicle inhalation. Animals were sacrificed after 30 min and pulmonary transcript levels of (A) IL-1β, (B) IL-6 or (C) TNF-α were assessed by real-time RT-PCR. Data were calculated relative to internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (−LPS, −MRS1754) ± SD (n=4–7). (D) Western-blot analysis of IL-6 protein assessed by Western blot. The same blot was stripped and re-probed for murine β-actin to control for loading conditions. One representative of three Western blots is displayed. Note enhanced lung inflammation following A2BAR antagonist treatment following LPS exposure.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Quantitative RT-PCR, Control, Western Blot

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: Endotoxin-induced acute lung injury was induced in 8–12 week old A2BAR−/− mice or littermate controls matched in age, gender and weight by inhalation of LPS over 30 min. Controls were treated with inhaled vehicle. Animals were sacrificed after 4h and pulmonary transcript levels of (A) IL-1β, (B) IL-6 or (C) TNF-α were assessed by real-time RT-PCR. Data were calculated relative to internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (-LPS) ± SD (n=5–13). (D) Western-blot analysis of IL-6 protein assessed by Western blot. The same blot was stripped and re-probed for murine β-actin to control for loading conditions. One representative of three Western blots is displayed. (E) Assessment of pulmonary edema by measurement of lung water content (n=5–6). (F) Measurement of pulmonary myeloperoxidase (MPO) as indicator for neutrophil accumulation (n=5–9). Note enhanced lung inflammation and pulmonary edema in A2BAR−/− mice following LPS exposure.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Quantitative RT-PCR, Control, Western Blot

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: Endotoxin-induced acute lung injury was induced in 8–12 week old A2BAR−/− mice or littermate controls matched in age, gender and weight by inhalation of LPS over 30 min. Controls were treated with inhaled vehicle. Animals were sacrificed, and pulmonary histology was assessed by a pathologist blinded to the treatment group. (A) Lung injury score (mean ± range, n=4). (B) Representative H&E staining (magnification 400x; one representative slide of 4). Note enhanced lung inflammation and tissue injury in A2BAR−/− mice exposed to LPS.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Staining

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: 8 to 12 week old A2BAR bone marrow chimeric mice (A2BAR+/+ → A2BAR+/+, WT → WT; A2BAR−/− → A2BAR+/+, WT → KO; A2BAR+/+ → A2BAR−/−, WT → KO and A2BAR−/− → A2BAR−/−, KO → KO) matched in age, gender and weight were exposed to 30 min of LPS or vehicle inhalation. Animals were sacrificed after 4h and (A) pulmonary transcript levels of IL-6 were determined by real-time RT-PCR. Data were calculated relative to internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (WT → WT treated with vehicle) ± SD (n=4–7).

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Quantitative RT-PCR, Control

Journal:

Article Title: Signaling through the A2B Adenosine Receptor Dampens Endotoxin-Induced Acute Lung Injury

doi: 10.4049/jimmunol.0903035

Figure Lengend Snippet: (A-C) 8–12 week old C57BL/6J mice matched in age, gender and weight were pre-treated with A2BAR agonist (BAY 60-6583, 2mg/kg i.p.) or vehicle, followed by 30 min of LPS or vehicle inhalation. Animals were sacrificed after 30 min and pulmonary myeloperoixidase (A), IL-6 protein levels measured by ELISA (B) or lung water content (C) were assessed (n=4). (D-E) Similar studies in gene-targeted mice for the A2BAR (A2BAR−/− mice; n=4). Note attenuated lung inflammation and pulmonary edema with A2BAR agonist treatment only in wild-type mice.

Article Snippet: Samples were incubated for 60 minutes with the following antibodies: Polyclonal rabbit anti-A2BAR antibody (Santa Cruz Biotechnology, Inc.) at a dilution of 1:1000 as primary antibody, rabbit Ig Fraction (Dako Cytomation, Glostrup, Denmark) as negative control.

Techniques: Enzyme-linked Immunosorbent Assay