Horseradish peroxidase hrp conjugated donkey anti rabbit immunoglobulin g igg

Dulbecco's modified Eagle's medium (DMEM), trypsin (0.25%), L-glutamine, penicillin/streptomycin, and fetal bovine serum (FBS) were purchased from Gibco (Gaithersburg, MD, USA). Andrographolide (≥98%), TNF-α, LY294002, SP600125, and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The anti-iNOS rabbit polyclonal antibody (pAb) and the anti-p65 antibody were purchased from Santa Cruz Biotechnology (Dallas, TX, USA); the anti-α-tubulin mouse monoclonal antibody (mAb) was purchased from Thermo Scientific (Waltham, MA, USA); and the anti-phospho-p38 MAPK Thr180/Tyr182 rabbit pAb, anti-p38 MAPK, anti-phospho-p44/p42 extracellular signal-regulated kinase (ERK1/2) Thr202/Tyr204 rabbit pAb, anti-ERK1/2 antibody, anti-phospho-JNK Thr183/Tyr185 rabbit mAb, anti-JNK antibody, anti-phospho-Akt Ser473 rabbit pAb, anti-Akt antibody, anti-phospho-p65 Ser536 rabbit pAb, and anti-IκBα antibody were purchased from Cell Signaling (Danvers, MA, USA). A hybond-P polyvinylidene difluoride (PVDF) membrane, an enhanced chemiluminescence (ECL) western blotting detection reagent and analysis system, the horseradish-peroxidase- (HRP-) conjugated donkey anti-rabbit immunoglobulin G (IgG), and the sheep anti-mouse IgG were acquired from Amersham (Buckinghamshire, UK). Andrographolide was dissolved in 0.1% DMSO and stored at 4°C until it was used.

Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), L-glutamine/penicillin/streptomycin, and anti-α-tubulin monoclonal antibodies (mAbs) were purchased from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA). LPS and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were bought from Sigma-Aldrich (St. Louis, MO, USA). Anti-iNOS, COX2, and Nrf2 polyclonal antibodies (pAbs) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). An anti-HO-1 pAb was purchased from Enzo (Farmingdale, New York, NY, USA). The anti-phospho-p65 (Ser536) mAb was purchased from Cell Signaling (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG) and sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, U.K.). Enhanced chemiluminescence (ECL) Western blotting detection reagent and Hybond-P polyvinylidene difluoride (PVDF) blotting membranes were purchased from GE Healthcare Life Sciences (Waukesha, WI, USA).

Auraptene (>98%), MG-132, and U73122 were purchased from Cayman Chem. (Ann Arbor, MI, USA). Luciferin/luciferase, collagen (type I), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), prostaglandin E₁ (PGE₁), heparin, 5,5-dimethyl-1 pyrroline N-oxide (DMPO), (E)-3-(4-methylphenylsulfonyl)-2-propenenitrile (BAY11-7082), Ro31-8220, and phorbol-12, 13-dibutyrate (PDBu) were purchased from Sigma (St Louis, MO, USA). Anti-IκBα (44D4) monoclonal antibody (mAb), anti-phospho-IκBα (Ser^32/36) (5A5), anti-NF-κB p65 mAbs, and anti-phospho-NF-κB p65 (Ser⁵³⁶) polyclonal antibody (pAb), anti-phospholipase C (PLC)γ2, anti-phospho (Tyr⁷⁵⁹) PLCγ2 pAbs were all from Cell Signaling (Beverly, MA, USA). An anti-pleckstrin (p47) pAb was purchased from Genetex (Irvine, CA, USA). The anti-α-tubulin mAb was from NeoMarkers (Fremont, CA, USA); and the Hybond-P polyvinylidene difluoride (PVDF) membrane, enhanced chemiluminescence (ECL) Western blotting detection reagent and analysis system, horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG), and sheep anti-mouse IgG were from Amersham (Buckinghamshire, UK). A 0.1% dimethyl sulfoxide (DMSO) solution was used to prepare the Auraptene suspension.

Collagen (type I) and thrombin were purchased from Sigma (St. Louis, MO, USA). The anti-phospho-c-Jun N-terminal kinase (JNK) (Thr¹⁸³/Tyr¹⁸⁵), anti-JNK, anti-phospho-p38 mitogen-activated protein kinase (MAPK), anti-p38 MAPK and anti-phospho-Syk (Tyr^525/526) monoclonal antibodies (mAbs) were purchased from Cell Signaling (Beverly, MA, USA). The anti-phospho-Akt (Ser⁴⁷³) and anti-Akt mAbs were purchased from Biovision (Mountain View, CA, USA). Anti-phospho-Fyn (Y⁵³⁰) mAb, anti-phospho-Lyn (Y⁵⁰⁷) mAb was obtained from Abcam (Cambridge, UK). The horseradish peroxidase (HRP) conjugated donkey anti-rabbit immunoglobulin G (IgG), the Hybond-P polyvinylidene difluoride (PVDF) membrane, the sheep anti-mouse IgG and the enhanced chemiluminescence western blotting detection reagent was purchased from Amersham (Buckinghamshire, UK). The TQ1, TQ-2 and TQ-3 were dissolved in DMSO and stored at 4 °C.

Collagen (type I), luciferin-luciferase, arachidonic acid (AA), U46619, PDBu, heparin, prostaglandin E1 (PGE1), 2,3,5-triphenyl tetrazolium chloride (TTC), and thrombin were purchased from Sigma (St. Louis, MO, USA). Fura 2-AM, an intracellular calcium indicator, was purchased from Molecular Probes (Eugene, OR, USA). The anti-phospho-p38 MAPK Ser182 monoclonal antibody (mAb) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-p38 MAPK, anti-phospho-JNK (Thr183/Tyr185), and anti-p44/42 ERK (1/2) mAbs and anti-PLCγ2, anti-phospho (Tyr759) PLCγ2, the anti-phospho-(Ser) PKC substrate, anti-JNK and anti-phospho-p44/p42 ERK (Thr202/Tyr204) polyclonal antibodies (pAbs) were purchased from Cell Signaling (Beverly, MA, USA). The anti-phospho-Akt (Ser473) and anti-Akt mAbs were purchased from Biovision (Mountain View, CA, USA). Anti-pleckstrin (p47) pAb was purchased from GeneTex (Irvine, CA, USA). The anti-α-tubulin mAb was purchased from NeoMarkers (Fremont, CA, USA). A hybond-P PVDF membrane, an enhanced chemiluminescence Western blotting detection reagent, the horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG), and the sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, UK). The Dade Behring PFA-100 CEPI and CADP test cartridges were obtained from Siemens Healthcare (Erlangen, Germany).

Amarogentin (88.9%) was purchased from ChromaDex (Irvine, CA). Arachidonic acid (AA), collagen (type I), 9,11-dideoxy-11α,9α-epoxymethanoprostaglandin (U46619), luciferin-luciferase, thrombin, SQ22536, phorbol-12,13-dibutyrate (PDBu), and 1H-[1,2,4]qxadiazolo[4,3-a]quinoxalin-1-one (ODQ) were purchased from Sigma (St. Louis, MO). The anti-phospho-c-Jun N-terminal kinase (JNK) (Thr¹⁸³/Tyr¹⁸⁵) and anti-phospho-p38 mitogen-activated protein kinase (MAPK) monoclonal antibodies (mAbs), and the anti-phospho-p44/p42 extracellular signal-regulated kinase (ERK) (Thr²⁰²/Tyr²⁰⁴), anti-phospholipase Cγ2 (PLCγ2), and anti-phospho (Tyr⁷⁵⁹) PLCγ2 polyclonal antibodies (pAbs) were purchased from Cell Signaling (Beverly, MA). The anti-phospho-p38 MAPK Ser¹⁸² mAb was purchased from Santa Cruz (Santa Cruz, CA). The anti-α-tubulin mAb was purchased from NeoMarkers (Fremont, CA). The anti-phospho-Akt (Ser⁴⁷³) and anti-Akt mAbs were purchased from Biovision (Mountain View, CA). The horseradish peroxidase- (HRP-) conjugated donkey anti-rabbit immunoglobulin G (IgG), the Hybond-P polyvinylidene difluoride (PVDF) membrane, the sheep anti-mouse IgG, and the enhanced chemiluminescence western blotting detection reagent were purchased from Amersham (Buckinghamshire, UK). The Amarogentin was dissolved in DMSO and stored at 4°C.