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2 protocols using anti phospho p38 mapk mab

1

Antibody Analysis of Cell Signaling Pathways

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The anti-caspase-9 and anti-caspase-3 monoclonal antibodies (mAbs), anti-Bax polyclonal antibody (pAb), anti-phospho-p53 pAb, anti-p53 mAb, anti-phospho-p38 MAPK mAb, anti-p38 MAPK mAb, and anti-phospho-SHP-1 mAb were purchased from Cell Signaling (Beverly, MA, USA). The anti-phospho-PP2A-C mAb, anti-demethylated-PP2A-C mAb, and anti-SHP-1 pAb were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The anti-PP2A-C pAb was purchased from Genetex (San Antonio, TX, USA), and the anti-α-tubulin mAb was purchased from NeoMarkers (Fremont, CA, USA). The horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG) and sheep anti-mouse IgG were purchased from Amersham.
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2

Murine Osteosarcoma Cell Culture

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The murine LM8 and Dunn OS cell lines were kindly donated by Dr Eugenie Kleinerman (M.D. Anderson Cancer Center, Houston, TX, USA). The cell lines were cultured in high-glucose Dulbecco’s modified eagle’s medium (DMEM-h; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin. The cultures were incubated at 37°C in a humidified atmosphere containing 5% CO2. Recombinant murine CXCL12 was purchased from PeproTech (Rocky Hill, NJ, USA) and AMD3100 from Merck (Darmstadt, Germany). The rabbit antibodies (Ab) used for western blotting were obtained from different resources: anti-CXCR4 polyclonal Ab, anti-CXCR7 polyclonal Ab (both from Abcam, Cambridge, MA USA), anti-phospho-SAPK/JNK monoclonal Ab (MAb) (Thr183/Tyr185), anti-SAPK/JNK MAb, anti-phospho-c-Jun MAb (Ser73), anti-c-Jun MAb, anti-phospho-Akt MAb (Ser473), anti-Akt MAb, anti-phospho-p38 MAPK MAb (Thr180/Tyr182), anti-p38 MAPK MAb, anti-phospho-p44/42 MAPK (Erk1/2; Thr202/Tyr204) MAb, anti-p44/42 MAPK (Erk1/2), anti-caspase-3 MAb, anti-caspase-8 MAb and anti-PARP MAb (all from Cell Signaling Technology, Inc., Danvers, MA, USA), and anti-β-actin (Santa Cruz Biotechnology, inc. Santa Cruz, CA, USA).
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