Reagents for sds page and western blot

Igepal CA-630, BSA, EDTA, PMSF, iodoacetamide, bicinchoninic acid, anti-FLAG M2–Peroxidase conjugate antibody, anti-HA-peroxidase conjugate and anti-β-Tubulin III antibody were from Sigma (St. Louis, MO). The anti-pERK1/2 and anti-ERK2 rabbit polyclonal IgGs were from Santa Cruz Biotechnology (Santa Cruz, CA). The synthetic αMSH analogue [Nle4, D-Phe7]-αMSH (NDP-MSH) and the protein synthesis inhibitor cycloheximide were from Calbiochem (Darmstadt, Germany). The radioligand [¹²⁵I]-NDP-MSH, specific activity 2000 Ci/mmol was from Amersham (Little Chalfont, Buckinghamshire, UK). The cAMP immunoassay kit was from Arbor Assays (Eisenhower Place, Michigan, USA). Lipofectamine 2000 was from Invitrogen (Carlsbad, CA). Reagents for SDS-PAGE and Western blot were from Bio-Rad (Richmond, CA, USA). Other reagents were from Merck (Darmstadt, Germany) or Prolabo (Barcelona, Spain).

Protease or phosphatase inhibitors and common laboratory reagents were from Sigma (St. Louis, MO, USA), Calbiochem (Darmstadt, Germany), Merck (Darmstadt, Germany) or Prolabo (Barcelona, Spain), unless specified otherwise. Lipofectamine 2000 was from Invitrogen. Opti-MEM I and other cell culture reagents were from Gibco (Gaithersburg, MD, USA). Reagents for SDS-PAGE and Western blot were from Bio-Rad (Richmond, CA, USA).