rabbit anti-human cpla2 polyclonal antibody  (Merck KGaA)

Journal: Biochimica et biophysica acta

Article Title: Effect of azithromycin on the LPS-induced production and secretion of phospholipase A2 in lung cells.

doi: 10.1016/j.bbadis.2015.03.008

Figure Lengend Snippet: Fig. 5. Effect of LPS and azithromycin (Az) on the production of cPLA2, as well as the production and secretion of sPLA2-IIA, from alveolar macrophages. Cells were isolated from four control and six ARDS patients. Incubation with LPS took place in the presence (5 and 20 μg/mL) or absence of azithromycin. Analysis took place by western blotting using 5 μg protein loading and appropriate antibodies for cPLA2, pcPLA2 and sPLA2-IIA. Antibodies against β-actin were used as control of protein loading. (A) Levels of cPLA2 and its activated form, pcPLA2. (B) Intracel- lular levels of sPLA2-IIA. (C) Levels of sPLA2-IIA in cell supernatants. Gel images are representative of one AM preparation from control and one from ARDS patient under different treatment conditions.

Article Snippet: The membranes were then incubated overnight with polyclonal rabbit anti-human sPLA2 group IIA (dilution 1:1000, group II sPLA2 (H-74): sc-20105, Santa Cruz Biotechnology, Inc. USA), polyclonal rabbit anti-human cPLA2 (dilution 1:1000, sc-454, Santa Cruz Biotechnology, Inc. USA) and polyclonal rabbit anti-human phosphorylated cPLA2 antibody (Ser 505) (dilution 1:1000) (#2831, Cell Signalling Technology, Beverly, MA, USA).

Techniques: Isolation, Control, Incubation, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Effect of azithromycin on the LPS-induced production and secretion of phospholipase A2 in lung cells.

doi: 10.1016/j.bbadis.2015.03.008

Figure Lengend Snippet: Fig. 5. Effect of LPS and azithromycin (Az) on the production of cPLA2, as well as the production and secretion of sPLA2-IIA, from alveolar macrophages. Cells were isolated from four control and six ARDS patients. Incubation with LPS took place in the presence (5 and 20 μg/mL) or absence of azithromycin. Analysis took place by western blotting using 5 μg protein loading and appropriate antibodies for cPLA2, pcPLA2 and sPLA2-IIA. Antibodies against β-actin were used as control of protein loading. (A) Levels of cPLA2 and its activated form, pcPLA2. (B) Intracel- lular levels of sPLA2-IIA. (C) Levels of sPLA2-IIA in cell supernatants. Gel images are representative of one AM preparation from control and one from ARDS patient under different treatment conditions.

Article Snippet: The membranes were then incubated overnight with polyclonal rabbit anti-human sPLA2 group IIA (dilution 1:1000, group II sPLA2 (H-74): sc-20105, Santa Cruz Biotechnology, Inc. USA), polyclonal rabbit anti-human cPLA2 (dilution 1:1000, sc-454, Santa Cruz Biotechnology, Inc. USA) and polyclonal rabbit anti-human phosphorylated cPLA2 antibody (Ser 505) (dilution 1:1000) (#2831, Cell Signalling Technology, Beverly, MA, USA).

Techniques: Isolation, Control, Incubation, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Impaired phospholipases A₂production by stimulated macrophages from patients with acute respiratory distress syndrome.

doi: 10.1016/j.bbadis.2010.06.008

Figure Lengend Snippet: Fig. 5. Immunoblot for cytosolic PLA2 (cPLA2) and its activated-phosphorylated form (p-cPLA2) in blood monocytes from control patients and those with primary or secondary ARDS. Cells from patients with primary and secondary ARDS and control patients were subjected to immunoblotting using anti-p-cPLA2 and anti-cPLA2 antibodies. Cells were treated with LPS or IFN-γ. Immunoblots for cPLA2 and p-cPLA2 were obtained by using polyclonal antibodies and equal protein loading was confirmed by anti-β-actin antibody. They are representative of three independent experiments. Monocytes from both control and primary ARDS patients responded to treatment by LPS and IFN-γ elevating the p-cPLA2 signal. In secondary ARDS, however, there was a persistent increase of p-cPLA2 levels, even in the non-treated cells, which did not change significantly after cell stimulation. The cPLA2 and p-cPLA2 bands appeared at 85 and 110 kDa, respectively, while β-actin at 43 kDa. (U): Untreated cells.

Article Snippet: The membraneswere then incubatedwith the appropriate antibody for 2 h: polyclonal rabbit anti-human cPLA2 (dilution 1:1000), polyclonal rabbit anti-human phosphorylated cPLA2 antibody (Ser 505) (dilution 1:1000), Cell Signalling Technology, (Beverly, MA, USA) or mouse anti-human sPLA2 group IIA.

Techniques: Western Blot, Control, Cell Stimulation

Journal: Biochimica et biophysica acta

Article Title: Impaired phospholipases A₂production by stimulated macrophages from patients with acute respiratory distress syndrome.

doi: 10.1016/j.bbadis.2010.06.008

Figure Lengend Snippet: Fig. 4. Immunoblot of cytosolic PLA2 (cPLA2) and its activated-phosphorylated form (p-cPLA2) in alveolar macrophages from patients with primary, secondary ARDS and from control patients. Alveolar macrophages isolated as described in Materials and methods were treated ex vivo with LPS (25 μg/mL) or IFN-γ (300 U/mL). (A): Immunoblots for cPLA2 and p-cPLA2 were obtained by using polyclonal antibodies and equal protein loading was confirmed by anti-β-actin antibody. They are representative of three independent experiments. Phosphorylation of cPLA2 was induced only in AMΦ from control patients after stimulation with both LPS and IFN-γ. The cPLA2 and p-cPLA2 bands appeared at 85 and 110 kDa, respectively, while β-actin at 43 kDa. (U): Untreated cells.

Article Snippet: The membraneswere then incubatedwith the appropriate antibody for 2 h: polyclonal rabbit anti-human cPLA2 (dilution 1:1000), polyclonal rabbit anti-human phosphorylated cPLA2 antibody (Ser 505) (dilution 1:1000), Cell Signalling Technology, (Beverly, MA, USA) or mouse anti-human sPLA2 group IIA.

Techniques: Western Blot, Control, Isolation, Ex Vivo, Phospho-proteomics