Anti jnk antibody

UTI was provided by Mochida Pharmaceutical Company. Antibody directed phosphorylated inhibitory-κBα (p-I-κB), p-ERK (1/2), p-MEK, p-JNK antibodies were obtained from Cell Signaling Biotechnology. Anti-I-κB, anti-NF-κB (p65 subunit), anti-ERK (1/2), anti-MEK, anti-JNK antibodies and anti-actin antibodies were purchased from Santa Cruz Biotechnology. LPS (200 pg/ml), MTT and other reagents were purchased from Sigma Chemical. All reagents were tested for endotoxin using the high-sensitivity version of the Limulus Amebocyte Lysate assay (Biowhittaker) and were from the lots containing the lowest amounts of endotoxin available.
Spherical PMMA particles (Polysciences) 1–10 μm in diameter (6.0 μm mean diameter, 95% < 10 μm). The particles were sterilized in 70% ethanol and incubated overnight with shaking at 4 °C in sterile PBS with penicillin (100 units/ml) and streptomycin (100 μg/ml) at a concentration of 0.5 mg/ml (2×10⁸ particles/25 μl) until use. Adherent endotoxin on the particles (620 EU/ml) was measured using the high-sensitivity version of the Limulus Amebocyte Lysate Assay (Biowhittaker) in the presence of a β-glucan blocking reagent (Biowhittaker).

Tumor necrosis factor-α (TNFα) was from PeproTech France (Neuilly-Sur-Seine, France). Apocynin (4-hydroxy-3-methoxyacetophenone), o-dianisidine hydrochloride, and thiobarbituric acid were obtained from Sigma Aldrich (Saint-Quentin Fallavier, France). Mouse KC/CXCL1 ELISA Kit was from RayBiotech, Inc. (Norcross, GA, USA). One-Step RT-PCR kit was from Qiagen S.A.S.-France (Courtaboeuf, France). Mouse anti-NOX1, -NOXA1, and -NOXO1 were a kind gift from Dr. R. Brandes (Instituts für Kardiovaskuläre Physiologie, Germany). In some experiments, rabbit polyclonal anti-NOX1 antibody from Sigma Aldrich (Saint-Quentin Fallavier, France), anti-NOXA1 antibody raised against the full-length protein from Sigma Aldrich (Saint-Quentin Fallavier, France), and anti-NOXO1 antibody directed against full-length proteins from Morphosys AbD GmbH (Düsseldorf, Germany) were also used. Anti-p22^PHOX and anti-DUOX2 (Y-15) were from Santa-Cruz Biotechnology (Heidelberg, Germany). Anti-phospho-ERK 1/2 and anti-phospho-p38MAPK antibodies were from R&D systems Europe (Abingdon, UK). Anti-phospho-JNK was from Cell Signaling Technology (Danvers, USA). Anti-ERK 1/2, anti-p38MAPK, and anti-JNK antibodies were from Santa-Cruz Biotechnology (Heidelberg, Germany).

H₂O₂ and catalase were purchased from Sigma. Antibodies against p-ERK1/2, ERK1/2, p-Src Y419, p-Src Y530 and Src were obtained from Cell Signaling (Beverly, MA, USA). Anti-Csk, anti-p-p38, anti-p38, anti-p-JNK, anti-JNK antibodies were obtained from Santa Cruz Technology (Santa Cruz, CA, USA). Antibody against caveolin-1 was obtained from BD Biosciences (San Diego, CA, USA). Tubulin antibody was obtained from Sigma. All Horseradish peroxidase (HRP)-conjugated secondary antibodies for immunoblotting were obtained from Santa Cruz Technology. Alexa flour 568-conjugated rabbit secondary antibody was obtained from Life Technologies.