Horseradish peroxidase conjugated anti phosphotyrosine py 20

Antibodies for total IGF1R (#3027), phosphorylated IGF1R tyrosine 1135 (#3918), phosphorylated AKT serine 473 (#9271) and threonine 308 (#9275), total p44/42 MAPK (#9102), phosphorylated p44/42 MAPK (#4376), phosphorylated p70 S6 kinase (#9205) used in immunoblotting were purchased from Cell Signaling Technology. Normal mouse IgG (sc-2025), InsR antibody for immunoprecipitation (sc-57342) and InsR antibody for immunoblotting (sc-711) were purchased from Santa Cruz Biotechnology. Horseradish peroxidase-conjugated anti-phosphotyrosine (pY-20) was purchased from BD Transduction Lab. (#610012). Anti-rabbit horseradish peroxidase-conjugated secondary antibody was purchased from Pierce.

Growth media and supplements were purchased from Invitrogen (Grand Island, NY). IGF-I was purchased from GroPep (Adelaide, Australia). IGF-II was purchased from Gemini (Woodland, CA). Insulin was purchased from Eli Lilly (Indianapolis, IN). LY294002, U0126, and actin antibody were purchased from Sigma-Aldrich (St. Louis, MO). Humanized anti-IGF-IR monoclonal antibody huEM164 was provided by Immunogen Inc. (Norwood, MA). BMS-754807 was provided by Bristol Myers Squibb (New Jersey, Marco Gottardis). Antibodies for phosphorylated AKT serine 473, total and phospho-IGF-IR, total and phosphorylated p44/42 (MAPK), phospho-p38 ^MAPK, total and phospho-IRS-1 (serine 636/639), phospho-S6K1, p21, p18, CDK2, CDK6, cyclin D1, cyclin D3, and PARP were purchased from Cell Signaling Technology (Beverly, MA). The IRS-2, total ERα, IGF-IRα, InRβ antibodies, and protein agarose A were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase-conjugated anti-phosphotyrosine (PY-20) was purchased from BD Biosciences (San Jose, CA). Anti-rabbit and anti-mouse horseradish peroxidase-conjugated secondary antibodies were purchased from Pierce (Rockford, IL). NT157 compound was kindly provided by A. Levitzki (The Hebrew University of Jerusalem, Israel).