Azoxymethane (AOM), 10 % (v/v) neutral buffered formalin, TRI Reagent®, and specific primers were bought from Sigma (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM), RPMI-1640 medium, Mycoplex™ fetal bovine serum (FBS), penicillin and streptomycin (100×), and trypsin EDTA (1×) were obtained from PAA Laboratories GmBH (Pasching, Austria). Colorimetric assay kit for caspase was bought from Genscript Corporation Inc (Piscataway, NJ, USA). High capacity RNA-to-cDNA Kit and SYBR® Select Master Mix (CFX) were purchased from Applied Biosystems (Foster City, CA, USA). Western blotting reagents were purchased from Bio-Rad (Hercules, CA, USA). All other chemicals and reagents used were of analytical grade and bought from Sigma-Aldrich (St. Louis, MO, USA).
Western blotting reagents
Manufactured by Bio-Rad
Sourced in United States
Western blotting reagents are a set of laboratory equipment and chemicals used to detect and analyze specific proteins within a complex sample. These reagents facilitate the separation, transfer, and visualization of proteins, allowing researchers to study their expression, interactions, and modifications.
Automatically generated - may contain errors
Lab products found in correlation
Acacetin, by Merck Group (2 mentions)
Tissue culture reagents and media, by Merck Group (2 mentions)
Western blotting lysis buffer, by Merck Group (2 mentions)
Secondary antibodies for western blotting, by Santa Cruz Biotechnology (2 mentions)
α napthoflavone, by Merck Group (2 mentions)
Dithiothreitol (dtt), by Merck Group (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Specific primers, by Merck Group (1 mentions)
Azoxymethane aom, by Merck Group (1 mentions)
Sybr select master mix, by Thermo Fisher Scientific (1 mentions)
High capacity rna to cdna kit, by Thermo Fisher Scientific (1 mentions)
Tri reagent, by Merck Group (1 mentions)
Juglone, by Merck Group (1 mentions)
Dextran t500, by GE Healthcare (1 mentions)
Hrp conjugated goat anti mouse, by Santa Cruz Biotechnology (1 mentions)
Hank s balanced salt solution, by Merck Group (1 mentions)
Ficoll, by GE Healthcare (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
9 protocols using western blotting reagents
1
Azoxymethane-induced Colorectal Cancer Model
2
Phospho-p47phox Signaling Pathway
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Lipopolysaccaride (LPS) from E. Coli O111:B4 strain, Juglone, PiB, Phosphate Buffered Saline (PBS), Hanks' Balanced Salt Solution (HBSS), protease and phosphatase inhibitors were obtained from Sigma Aldrich (Saint Quentin Fallavier, France). Dextran T500 and Ficoll was from GE healthcare (Orsay, France). Sodium dodecyl-sulfate polyacrylamide (SDS-PAGE) and western blotting reagents were supplied by Bio-Rad (Hercules, CA, USA). The rabbit polyclonal antibodies against phospho-p47phox sites (phospho-Ser345, phospho-Ser320, phospho-Ser304, phospho-Ser315, phospho-Ser328), p67phox, and p47phox were produced by our lab as described elsewhere (18 (link), 33 (link)). Anti-phospho(P)-ERK1/2, ERK1/2, P-p38, and p38 were from cell signaling Technology (Boston, MA, USA). HRP-conjugated goat anti-mouse were from Santa Cruz Biotechnology Inc. (Heidelberg, Germany).
3
Western Blot Analysis of Apoptosis Regulators
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Chemicals and solvents, radioimmunoprecipitation assay (RIPA) lysis buffer (cat. no. R0278) were used without further purification and were supplied by Sigma-Aldrich; Merck KGaA (Darmstadt, Germany), unless otherwise stated. Media and cell culture reagents were from Biological Industries (Kibbutz Beit Haemek, Israel). Western blotting reagents were obtained from Bio-Rad Laboratories Inc. (Hercules, CA, USA). DAPI and primary antibodies against poly(ADP-ribose) polymerase 1/2 (PARP1/2; cat. no. sc-7150), p53 (cat. no. sc-126), p21 (cat. no. sc-756), BCL2 associated X protein (BAX; cat. no. sc-7480), cyclin B (cat. no. sc-sc-53236), AKT (cat. no. sc-514032) and tubulin (cat. no. sc-5286) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Primary antibodies against B-cell lymphoma 2 (BCL2; cat. no. 2876) and caspase-9 (cat. no. 9502) were provided by Cell Signaling Technology, Inc. (Danvers, MA, USA). Horseradish peroxidase-conjugated secondary antibodies were provided by Bio-Rad Laboratories, Inc. (Hercules, CA, USA) and electrochemiluminescence reaction (ECL) detection system was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA).
4
Comprehensive Antibody and Reagent Sources
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All peptides were synthesized by CPC Scientific (Sunnyvale, CA). The SHLP antibodies were custom synthesized by Harlan (Indianapolis, IN) and YenZym (San Francisco, CA); all other antibodies were purchased from Cell Signaling Technology (Danvers, MA). Western blotting reagents were purchased from Biorad (Hercules, CA). Cell culture reagents, primary cortical neurons, dihydroethidine (DHE), TRIzol®, Oil Red-O, and calcein-AM were purchased from Life Technologies (Grand Island, NY). Antibodies against phospho-STAT3 (Y705), total STAT3, phospho ERK-1/2 (T202/Y204), and total ERK-1/2 were purchased from Cell Signaling Technology. LINCOplex™ was purchased from Millipore (Billerica, MA). All primers and chemicals were purchased from Sigma (St Louis, MO). Luminescent ATP assay kits were purchased from Promega (Madison, WI).
5
Inhibition of Cell Cycle Regulators
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The reagents for chemical synthesis were from Merck KGa (Darmstadt, Germany) and Thermo Fisher Scientific, Inc., (Waltham, MA, USA). Media and cell culture reagents were from Merck KGa. Western blotting reagents were obtained from Bio-Rad Laboratories Ltd., (Watford, Hertfordshire, UK). Primary antibodies for cyclin D1, p53, p21 and NF-κB were from Santa Cruz Biotechnology, Inc., (Dallas, TX, USA), whereas primary antibodies for JNK and p-JKNThr183/Tyr185 were from Upstate Biotechnology, Inc. (Lake Placid, NY, USA) and for β-actin from Merck KGa. Secondary antibodies used were from Santa Cruz Biotechnology, Inc.
6
Immunoblotting Protein Expression Analysis
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Immunoblotting was performed as previously described.[27 ] Briefly, cells or tumor tissues were lysed in RIPA buffer, and protein extraction was carried out on ice. The protein concentration was determined by a BCA protein assay kit (Boster Biological Technology). Total protein lysate was mixed with SDS loading buffer, and proteins were separated by electrophoresis on 10% SDS‐PAGE. After that, proteins were transferred onto PVDF membranes. After blocking with 5% skim milk in TBST for 2 h, the membranes were washed and incubated sequentially with primary antibody and anti‐mouse or anti‐rabbit secondary antibody. Immunostaining with the antibody was performed using ECL western blotting reagents, and densitometric analysis of the immunoblot was carried out using Image Lab software (Bio‐Rad).
7
Macrophage Gene and Protein Expression Analysis
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p-Cymene (99%) was purchased from Sigma-Aldrich Chemical Co (St. Louis, MO). RPMI Medium 1640(1X) mediums, FBS (Fetal Bovine Serum), and Pen Strep (Penicillin Streptomycin) were all purchased from Life Technologies (Carlsbad, CA). The murine macrophages were RAW 264.7 and purchased from ATCC (Manassas, VA). TRIzol Reagent was purchased from ThermoFisher Scientific (Waltham, MA). All the Gene expression reagents and Western Blotting reagents were obtained from Bio-Rad (Hercules, CA).
8
CYP1A1 and CYP1B1 Protein Expression
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), 7-ethoxyresorufin, resorufin, α-napthoflavone, acacetin, tissue culture reagents and media, Western blotting lysis buffer and DTT were purchased from Sigma Aldrich (St Louis, MO, USA). Tangeretin and 4ʹ OH tangeretin were purchased from Apin chemicals (Abingdon, UK). Western blotting reagents were from Bio-Rad (Berkeley, CA, USA). The polyclonal antibody for CYP1A1 was from Daiichi Pure Chemicals (Gentest corporation, MA, USA), whereas the monoclonal antibodies for CYP1B1 from Auvation Limited (Glasgow, Scotland, UK), and for βactin from Cell signaling (Leiden, Netherlands). Secondary antibodies for western blotting were from Santa Cruz Biotechnology (Dallas, TX, USA).
9
MTT assay and Western blotting protocol
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3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), propidium iodide (PI), α-napthoflavone, acacetin, apigenin, luteolin, kaempferol, quercetin, tissue culture reagents and media, Western blotting lysis buffer and DTT were purchased from Sigma Aldrich (St Louis, MO, USA). Scutellarein was purchased from Extrasynthese (Lyon, France) and 6-OH luteolin was synthesized as described in previous methodologies (Androutsopoulos et al., 2009c) . Western blotting reagents were from Bio-rad (Berkeley, CA, USA). The polyclonal antibody for CYP1B1 was from Gentest (BD Biosciences, CA, USA), whereas the monoclonal for β-actin from Cell signaling (Leiden, Netherlands). Secondary antibodies for western blotting were from Santa Cruz Biotechnology (Dallas, TX, USA).
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