Thiazolyl blue tetrazolium bromide (MTT), 2″,7″-dichlorodihydro-fluorescein diacetate (DCFH-DA) and other reagents were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Tris was purchased from Duchefa Biochemie (BH Haarlem, The Netherlands), dimethyl sulfoxide (DMSO) from Junsei Chemical Co. (Tokyo, Japan). Fetal bovine serum (FBS) and penicillin-streptomycin (P/S) were purchased from Gibco (Los Angeles, CA, USA). Dulbecco's Modified Eagle's Medium (DMEM) was purchased from Welgene Inc. (Gyeongsangbuk-do, Korea). BCA reagent and albumin standard were from Thermo Scientific (Waltham, MA, USA). Primary Bcl-2 antibody was purchased from Oncogene (Bracknell, England). Bax and PAPR antibodies were obtained from Cell Signaling (Danvers, MA, USA) and cytochrome c, caspase-9, and β-actin antibodies from Santa Cruz (Paso Robles, CA, USA). Caspase-3 antibody was obtained from EMD Millipore (Billerica, MA, USA). Secondary antibodies goat anti-rabbit IgG, pAb, goat anti-mouse IgG, pAb, and caspase-3 activity assay kit were obtained from Enzo Life Sciences (Farmingdale, NY, USA). West-Q chemiluminescent substrate was purchased from GenDEPOT (Katy, TX, USA). The solvents used were of HPLC grade, unless stated otherwise.
Goat anti mouse igg
Manufactured by Enzo Life Sciences
Sourced in United States
Goat anti-mouse IgG is a secondary antibody used in a variety of immunoassay techniques. It is designed to detect and bind to mouse immunoglobulin G (IgG) antibodies, allowing for their visualization or quantification.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit igg, by Enzo Life Sciences (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Hrp conjugated goat anti rabbit igg, by Enzo Life Sciences (2 mentions)
P akt, by Cell Signaling Technology (2 mentions)
P mtor, by Cell Signaling Technology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Bca assay kit, by Thermo Fisher Scientific (2 mentions)
Pro prep protein extraction buffer, by iNtRON Biotechnology (2 mentions)
Tnf α, by Thermo Fisher Scientific (2 mentions)
Cm h2dcfda, by Thermo Fisher Scientific (2 mentions)
Albumin standard, by Thermo Fisher Scientific (1 mentions)
Caspase 3 activity assay kit, by Enzo Life Sciences (1 mentions)
Bca reagent, by Thermo Fisher Scientific (1 mentions)
West q chemiluminescent substrate, by GenDEPOT (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Welgene (1 mentions)
Dimethyl sulfoxide (dmso), by Junsei (1 mentions)
Chemiluminescent detection, by Merck Group (1 mentions)
P p70s6k, by Cell Signaling Technology (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
6 protocols using goat anti mouse igg
1
Biochemical Assays for Cellular Cytotoxicity Analysis
2
Western Blot Analysis of AKT-mTOR Pathway
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Proteins were extracted from cell lysates using PRO-PREP protein extraction buffer (Intron biotechnology, Seoul, Korea) and protein concentration was determined by BCA assay kit (Thermo scientific, Rockford, IL, USA). Protein samples were separated with SDS-PAGE, and transferred to PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% skim milk and incubated with primary antibodies p-AKT (Ser 473), AKT, p-mTOR (Ser 2448), mTOR, p-P70S6K (Ser 371), P70S6K (Cell Signaling technology, Boston, MA, USA), TSC2, TBDC17, Rheb, beta actin from (Santa Cruz Biotechnology, Dallas, Texas, USA) overnight at 4 °C, followed by incubation with HRP conjugated goat anti-rabbit IgG (Enzo Life Sciences, Farmingdale, New York, NY, USA) and goat anti-mouse IgG (Enzo Life Sciences, Farmingdale, NY, USA) secondary antibody at 25 °C for 1 h and Western blots were enhanced using chemiluminescent detection (Immobilon, Millipore, Burlington, MA, USA).
3
Evaluation of Apoptosis and Autophagy
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Acridine orange (AO), 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA), 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI), N-acetyl-l -cysteine (NAC) and 3-methyladenine (3-MA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). A FITC Annexin V Apoptosis Detection Kit (BD Biosciences, San Jose, CA, USA) was obtained. A Caspase-3 colorimetric assay kit was obtained from R&D Systems Inc. (Minneapolis, MN, USA). LY294002, PD98059 and SB203580 were purchased from TOCRIS (Bristol, UK). The ECL Western Kit was purchased from Amersham (Arlington Heights, IL, USA). Antibodies against LC-3B, Beclin-1, β-actin, phosphor-AKT, total AKT, phosphor-ERK, total ERK, phospho-p38, total p38, phosphor-JNK, and total JNK were purchased from Cell Signaling Technology (Beverly, MS, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA). The goat anti-mouse IgG and goat anti-rabbit secondary antibodies were purchased from Enzo Life Science (Farmingdale, NY, USA). The FITC-conjugated anti-rabbit IgG secondary antibody was purchased from BioFX Laboratories (Owings Mills, MD, USA).
4
Phytochemical Analysis and Anti-inflammatory Effects
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Ferulic acid and 5-hydroxymethyl-2-furaldehyde (5-HMF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Albiflorin and paeoniflorin were the products of Wako (Osaka, Japan). Nodakenin was purchased from NPC BioTechnology Inc. (Daejeon, Korea). The purity of all reference standards was ≥98.0%. HPLC-grade methanol, acetonitrile, and water were obtained from J.T.Baker (Phillipsburg, NJ, USA). Glacial acetic acid, analytical reagent grade, was purchased from Junsei (Tokyo, Japan). RPMI 1640, fetal bovine serum, TNF-α, tissue culture reagents, 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxy-methylester (BCECF-AM), DAF-FM diacetate, and CM-H2DCFDA, Alexa Fluor 488 and 594 conjugated second antibodies were purchased from Invitrogen (San Diego, CA). Biotin 3′ End DNA Labeling Kit, LightShift® Chemiluminescent EMSA Kit, Biodyne® Precut Nylon Membranes, Lipofectamine LTX reagent, and Renilla-Firefly Luciferase Dual Assay Kit were purchased from Pierce Biotechnology (Rockford, USA). Primary antibodies, including mouse anti-ICAM-1, goat anti-VCAM-1, rabbit anti-E-selectin, mouse anti-NF-κB, mouse anti-p-IκB-α, rabbit anti-HO-1, and rabbit anti-Nrf2, were purchased from Santa Cruz Biotechnology (CA, USA). Donkey anti-goat IgG-H+I were purchased from Bethyl (Montgomery, USA) and goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Enzo (Farmingdale, USA).
5
Oxidative Stress Cell Signaling Pathway
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DMEM low glucose, fetal bovine serum, TNF-α, cell culture reagents, and CM-H2DCFDA were purchased from Invitrogen (San Diego, CA). Primary antibodies, including mouse anti-cyclin D1, rabbit anti-CDK4, mouse anti-cyclin E, rabbit anti-CDK2, rabbit anti-p21, mouse anti-p27, rabbit anti-MMP2, and rabbit anti-MMP9, were purchased from Santa Cruz Biotechnology (CA, USA). Goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Enzo (Farmingdale, USA).
6
Protein Extraction and Western Blot Analysis
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Total cell lysates were extracted using PRO-PREP protein extraction buffer (Intron biotechnology, Seoul, Korea), and protein concentration was determined by a BCA assay kit (Thermo Scientific, Rockford, IL, USA). Protein samples were separated with SDS-PAGE and transferred onto PVDF membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 5% skim milk for 1 h and incubated with primary antibodies, active beta-catenin, p-AKT (Ser 473, 308), AKT, p-mTOR (Ser 2448), mTOR, Raptor, Rictor (Cell Signaling Technology, Boston, MA, USA), p-TSC2,TSC2, p-GSK-3β, GSK-3β, LAMP1, LAMP2, Rheb, tubulin, Beta-Actin (from Santa Cruz Biotechnology, Dallas, Texas USA) and LC-3B (abcam) overnight at 4 °C, followed by incubation with HRP conjugated goat anti-rabbit IgG (Enzo Life Sciences, NY, USA) and goat anti-mouse IgG (Enzo Life Sciences, NY, USA) secondary antibodies at 25 °C for 1 h. Western blots used chemiluminescent detection (Immobilon, Millipore, Burlington, MA, USA).
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