Rabbit polyclonal antisera to atrogin 1 mafbx

The chymotrypsin substrate (N-Succinyl-Leu-Leu-Val-Tyr-7-Amido-4-methylcoumarin) was obtained from Sigma (Dorset, UK). L-[2,6-3 H] phenylalanine (sp.act.3.7 TBqmmol -1 ) was from Perkin Elmer (Cambridge, UK). Mouse monoclonal antibodies to 20S proteasome α-subunits and p42 were from Affiniti Research Products (Exeter, UK). Fatty acid free BSA, tumour necrosis factor (TNF-α), polyclonal rabbit antibody to mouse β-actin, chymotrypsin substrate (N-Succinyl-Leu-Leu-Val-Tyr-7-Amido-4-methylcoumarin) and lactacystin were purchased from Sigma-Aldridge (Dorset, UK). Peroxidase-conjugated sheep antirabbit and antimouse antibodies were purchased from GE Healthcare (Bucks, UK), as were Hybond A nitrocellulose membranes. Enhanced chemiluminescence (ECL) development kits were from Pierce through Thermo Fischer Scientific (Northumberland, UK). Rabbit polyclonal antisera to atrogin-1/MAFbx, and goat polyclonal antisera to MuRF1 were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). EPA was added to fatty acid free BSA in doubled distilled water (g/g) and neutralised with equimolar NaHCO 3 . Green tea extract (Sunphenon 90D) was purchased from Taiyo international, (Tokyo, Japan). Curcumin C3 was from Sabinsa Corporation, NJ, USA. EPA (free acid) (98> %pure) was from Enzo Life Sciences (Exeter, UK).