Horseradish peroxidase hrp conjugated donkey antirabbit igg

Licochalcone A (LA, ≥95%) was purchased from Cayman Chemical (Ann Arbor, MI, USA). Collagen, thrombin, and U46619 were purchased from Chrono-Log (Havertown, PA, USA). FITC-conjugated anti-P-selectin and PAC-1 antibodies were purchased from Biolegend (San Diego, CA, USA). FITC-conjugated Collagen, phorbol-12, 13-dibutyrate (PDBu), luciferase/luciferin, fluorescein sodium, and 2,3,5-triphenyltetrazolium chloride (TTC) were purchased from Sigma (St. Louis, MO, USA). Fura 2-AM was purchased from Molecular Probe (Eugene, OR, USA). Anti-phospho PLCγ2 (Tyr⁷⁵⁹), anti-PLCγ2, anti-phospho-(Ser) PKC substrate, anti-phospho-p38 MAPK (Ser¹⁸⁰/Tyr¹⁸²), anti-phospho-p44/42 MAPK (ERK1/2; Thr²⁰²/Tyr²⁰⁴), anti-c-Jun N-terminal kinase (JNK), and anti-phospho-Akt (Ser⁴⁷³) polyclonal antibodies and anti-p38 MAPK, anti-p44/42 MAPK, anti-phospho JNK (Thr¹⁸³/Tyr¹⁸⁵), and anti-Akt monoclonal antibodies were purchased from Cell Signaling (Beverly, MA, USA). The pleckstrin (p47) antibody was purchased from GeneTex (Irvine, CA, USA). The Hybond-P polyvinylidene difluoride membrane, an enhanced chemiluminescence (ECL) Western blotting detection reagent and analysis system, horseradish peroxidase (HRP)-conjugated donkey antirabbit IgG, and sheep antimouse IgG were purchased from Amersham (Buckinghamshire, UK). LA was dissolved in dimethyl sulfoxide (DMSO) and stored at 4 °C until use.

Type I collagen and phorbol-12, 13-dibutyrate (PDBu) were purchased from Sigma (St Louis, MO). Fura 2-AM was purchased from Molecular Probe (Eugene, OR). The anti-Akt (pan) (40D4) monoclonal antibody (mAb), anti-phospho-Akt (Ser⁴⁷³) polyclonal antibody (pAb), anti-phospho-(Ser) protein kinase C (PKC) substrate pAb, anti-phospho-p38 mitogen-activated protein kinase (MAPK) (Thr¹⁸⁰/Tyr¹⁸²) pAb, anti-p38 MAPK (5F11) mAb, anti-phospho-p44/42 MAPK (ERK1/2) (Thr²⁰²/Tyr²⁰⁴) pAb, anti-p44/42 MAPK (137F5) mAb, anti-phospho-c-Jun N-terminal kinse (JNK) (Thr¹⁸³/Tyr¹⁸⁵) mAb, and anti-JNK pAb were purchased from Cell Signaling (Beverly, MA). The anti-α-tubulin mouse mAb was purchased from Thermo Scientific (Waltham, MA). The Hybond-P polyvinylidene difluoride (PVDF) membrane, an enhanced chemiluminescence (ECL) western blotting detection reagent, a horseradish-peroxidase (HRP)-conjugated donkey anti-rabbit IgG, and a sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, UK).