rabbit polyclonal antibody against p38mapk  (Proteintech)

Journal: Journal of translational medicine

Article Title: Bax/Mcl-1 balance affects neutrophil survival in intermittent hypoxia and obstructive sleep apnea: effects of p38MAPK and ERK1/2 signaling.

doi: 10.1186/1479-5876-10-211

Figure Lengend Snippet: Figure 3 Effects of ERK and p38MAPK inhibition on Mcl-1 expression in various oxygen conditions. Neutrophils were incubated for 6 hrs at normoxia (Nox), sustained hypoxia (SH), or 6 cycles of intermittent hypoxia (IH) without or with 10 μM U0126 (a specific inhibitor of MEK1/2, which blocks ERK1/2 activation), and 30 μM SB202190 (a competitive inhibitor of the p38 kinase). Mcl-1 expression was assessed by confocal laser scanning microscopy. (A-I) Representative photomicrographs of Mcl-1 expression (green) from 1 out of 4 experiments performed. (A) Neutrophils incubated in Nox, (B) IH and (C) SH. (D) Neutrophils incubated in Nox with MEK1/2 inhibitor, (E) in IH with MEK1/2 inhibitor, and (F) in SH with MEK1/2 inhibitor. (G) Neutrophils incubated in Nox with p38MAPK inhibitor (H) in IH with p38MAPK inhibitor, and (I) in SH with p38MAPK inhibitor. (J) The average values of Mcl-1 expression for ERK1/2 inhibitor and (K) for p38MAPK inhibitor by relative percent. The intensity of Mcl-1 expression at normoxia without inhibitors was considered as 100%, and the changes induced by the IH and SH with or without the inhibitors were plotted as a relative percentage of this value. P values represent significance of untreated vs. inhibitor treated neutrophils in each oxygen condition.

Article Snippet: Membranes were blocked and incubated with primary rabbit polyclonal antibodies against Thr180/Tyr182-phosphorylated p38MAPK (Cell Signaling Technology, Inc., Beverly, MA, USA), Thr202/Tyr204-phosphorylated ERK1/2 (Cell Signaling Technology, Inc., Beverly, MA, USA), Bax (N20, sc493) and Mcl-1 (S-19, sc819, Santa Cruz Biotechnology Inc., CA, USA), followed by goat anti-rabbit IgG incubation (Amersham Biosciences Europe GmbH, Freiburg, Germany).

Techniques: Inhibition, Expressing, Incubation, Activation Assay, Confocal Laser Scanning Microscopy

Journal: Journal of translational medicine

Article Title: Bax/Mcl-1 balance affects neutrophil survival in intermittent hypoxia and obstructive sleep apnea: effects of p38MAPK and ERK1/2 signaling.

doi: 10.1186/1479-5876-10-211

Figure Lengend Snippet: Figure 4 ERK1/2 and p38MAPK phosphorylation in neutrophils exposed to various oxygen conditions. (A) The average (± SD) normalized values of p-ERK1/2 (n=6), obtained by densitometric analysis of western blots, are presented as fold increase over β-actin in arbitrary units in normoxia (Nox), intermittent hypoxia (IH) and sustained hypoxia (SH). (B) A representative western blot depicts the dual phosphorylated (Thr202/Tyr204) form of ERK1/2 (p-ERK1/2) and dual phosphorylated (Thr180/Tyr182) form of p38MAPK (p-p38) compared to non-phosphorylated ERK1/2 and p38MAPK in Nox, IH, and SH.

Article Snippet: Membranes were blocked and incubated with primary rabbit polyclonal antibodies against Thr180/Tyr182-phosphorylated p38MAPK (Cell Signaling Technology, Inc., Beverly, MA, USA), Thr202/Tyr204-phosphorylated ERK1/2 (Cell Signaling Technology, Inc., Beverly, MA, USA), Bax (N20, sc493) and Mcl-1 (S-19, sc819, Santa Cruz Biotechnology Inc., CA, USA), followed by goat anti-rabbit IgG incubation (Amersham Biosciences Europe GmbH, Freiburg, Germany).

Techniques: Phospho-proteomics, Western Blot

Journal: Molecular Vision

Article Title: Protective effect of paeoniflorin against oxidative stress in human retinal pigment epithelium in vitro

doi:

Figure Lengend Snippet: Mitogen-activated protein kinase (MAPK) were activated by H 2 O 2 in ARPE-19 cells. A : Human RPE cells were exposed to 200 μM H 2 O 2 for various time points (30, 60, 120 min) and cell lysates were prepared. The phosphorylated and total protein levels of p38 mitogen-activated protein kinase (p38MAPK), extracellular signal regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) were detected with specific antibodies, using western blot analysis as described in the Methods. β-actin served as the loading control. Figures were selected as representative data from three independent experiments. B : After 24-h exposure to H 2 O 2 , cell viability was measured with an MTT assay. Each value represents the mean±SEM of three independent experiments (n=3 experiments, *p<0.05).

Article Snippet: After blocking nonspecific binding with 5% skim milk, the membranes were incubated with a rabbit polyclonal antibody against p38MAPK (1:1,000), ERK1/2 (1:1,000), JNK (1:1,000), phosphorylated forms of p38MAPK (1:1,000), ERK1/2 (1:2,000) or JNK (1:1,000; Cell Signaling Technology, Danvers, MA), followed by incubation with a horseradish peroxidase–conjugated goat antibody against rabbit immunoglobulin (IgG; 1:2,000; Cell Signaling Technology, Danvers, MA).

Techniques: Western Blot, Control, MTT Assay

Journal: Molecular Vision

Article Title: Protective effect of paeoniflorin against oxidative stress in human retinal pigment epithelium in vitro

doi:

Figure Lengend Snippet: Paeoniflorin (PF) inhibits H 2 O 2 -induced MAPK activation in ARPE-19 cells. ARPE-19 cells were pretreated with different concentrations of PF for 4 h and exposed to 200 μM H 2 O 2 for 30 min; cell lysates were then prepared. The phosphorylated and total protein levels of p38MAPK and ERK1/2 were detected with specific antibodies, using western blot analysis as described in the Methods. β-actin served as the loading control. Figures were selected as representative data from three independent experiments. Quantitative analysis was performed by measuring the intensity relative to the control. Each value represents the mean±SEM of three independent experiments (n=3 experiments,*p<0.05).

Article Snippet: After blocking nonspecific binding with 5% skim milk, the membranes were incubated with a rabbit polyclonal antibody against p38MAPK (1:1,000), ERK1/2 (1:1,000), JNK (1:1,000), phosphorylated forms of p38MAPK (1:1,000), ERK1/2 (1:2,000) or JNK (1:1,000; Cell Signaling Technology, Danvers, MA), followed by incubation with a horseradish peroxidase–conjugated goat antibody against rabbit immunoglobulin (IgG; 1:2,000; Cell Signaling Technology, Danvers, MA).

Techniques: Activation Assay, Western Blot, Control