Goat anti rabbit igg horseradish peroxidase hrp sc 2004

Immunoblot was done as described before [21 (link)]. Rabbit anti-Actin antibody (A5060, 1:15000 dilution) was from Sigma-Aldrich (Saint Louis, Missouri, USA). Rabbit antibody against human BIK protein (#4592) and TCF3/E2A (#4865) was from Cell Signaling (1:1000 dilution). Secondary antibodies included goat anti-rabbit IgG-horseradish peroxidase (HRP; sc-2004), donkey anti-goat IgG-horseradish peroxidase (HRP; sc-2020; both Santa Cruz Biotechnology). Immunoblots were quantified using ImageJ software. Anti-ACTB antibody was used as a loading control. The relative expression levels of target proteins were normalized to those of ACTB proteins.

Garlic extract was obtained from Egarak (Busan, Korea). Polyclonal antibodies against p-Cdc2 p34 (sc-12340-R), Cdc2 p34 (sc-54), CHK2 (sc-9064), Cdc25c (sc-327), p-Cdc25c (sc12354), p21WAF1 (sc-756), p53 (sc-126), Cyclin A (sc-751), Cyclin B1 (sc-245), p-ATM (sc-47739), ATM (sc-23921), WEE1 (sc-325), HSPA6 (sc-374589) and GAPDH (sc-20357) were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Polyclonal antibodies against CHK1 (2360), p-CHK1 (2341), p-CHK2 (2661), ERK (9102), p-ERK (9101), JNK (9258), p-JNK (9251), p38 MAP kinase (9212), p-p38 MAP kinase (9211), AKT (9272), and p-AKT (9271) were obtained from Cell Signaling Technology Inc. (Danvers, MA, USA). Goat anti-rabbit IgG-horseradish peroxidase (HRP) (sc-2004), goat anti-mouse IgG-HRP (sc-2005), and donkey anti-goat IgG-HRP (sc-2020) were purchased from Santa Cruz Biotechnology Inc. Western Lightning Plus-ECL was obtained from PerkinElmer, Inc. (PerkinElmer, MA, USA). U0126, SB203580, SP600125, and wortmannin, were obtained from Calbiochem (San Diego, CA). A Nuclear Extract kit and EMSA Gel Shift kit were obtained from Panomics (Fremont, CA, USA). HSPA6 cDNA was obtained from the Korea human gene bank.

(±)-L-Alliin and S-allylcysteine (SAC) was purchased from Sigma-Aldrich (St. Louis, MO., USA). LPS (purified from Escherichia coli (E. coli) O127: E8), penicillin-G, streptomycin, sulfanilamide, N-(1-naphtyl)ethylenediamine dihydrochloride, and 2’,7’-dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO., USA). IL-6 and TNF-α Enzyme Linked Immuno Sorbent Assay (ELISA) kit was purchased from Enzo Life Science Inc. (Farmingdale, NY, USA). The primary antibodies against iNOS (#2982), COX-2 (#4842), NF-κB (p65) (#8242) and IκBα (#4812) were obtained from Cell Signaling (Cell Signaling Tech., Beverly, MA, USA). Goat anti-rabbit IgG-horseradish peroxidase (HRP) (sc-2004) and goat anti-mouse IgG-HRP (sc-2005) were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and dimethylsulfoxide (DMSO) were obtained from Hyclone Laboratories (Logan, UT, USA).