Western blotting analysis was performed as described previously [30 (link)]. Primary antibodies against LSH and α-tubulin were purchased from Santa Cruz Biotechnology, and primary antibody against GINS4 was purchased from GeneTex. EMT Antibody Sampler Kit and primary antibodies against histone H3 were purchased from Cell Signaling Technology, and primary antibody against β-actin was purchased from Sigma-Aldrich (St. Louis, MO).
Primary antibody against β actin
Manufactured by Merck Group
Sourced in United States
The primary antibody against β-actin is a laboratory reagent used to detect and quantify the presence of the β-actin protein in biological samples. β-actin is a highly conserved cytoskeletal protein found in all eukaryotic cells and is commonly used as a loading control or reference protein in various biochemical and cell biology experiments.
Automatically generated - may contain errors
Lab products found in correlation
Ribonuclease a, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
Emt antibody sampler kit, by Cell Signaling Technology (1 mentions)
Ketamine, by Merck Group (1 mentions)
Hematoxylin and eosin, by Merck Group (1 mentions)
Salidroside, by Merck Group (1 mentions)
Enhanced chemiluminescence kit, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase labelled secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Protein quantification kit, by Bio-Rad (1 mentions)
Malondialdehyde analyzing kit, by Nanjing Jiancheng (1 mentions)
Matrigel growth factor reduced gfr basement membrane matrix, by Corning (1 mentions)
Tritc conjugated phalloidin, by Cell Signaling Technology (1 mentions)
Ham s f 12 nutrient mix, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Lab tek 2 8 well chamber slide, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Bio-Rad (1 mentions)
Signalfire ecl reagent, by Cell Signaling Technology (1 mentions)
Hoechst 33342, by Cell Signaling Technology (1 mentions)
Bradford solution, by Bio-Rad (1 mentions)
10 protocols using primary antibody against β actin
1
Western Blotting of EMT Markers
2
Oxidative Stress Biomarkers Analysis
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Salidroside, ketamine, primary antibody against β-actin, and hematoxylin and eosin were supplied by Sigma Chemical Company (St. Louis, MO, USA). A malondialdehyde analyzing kit was obtained from Nanjing Jiancheng Institute of Bioengineering (Nanjing City, China). Protein quantification kit was purchased from Bio-Rad Laboratories (Hercules, CA, USA). The primary antibodies against copper- and zinc-containing (Cu-Zn) superoxide dismutase and catalase, horseradish peroxidase-labelled secondary antibody, and an enhanced chemiluminescence kit were provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA). The other chemicals used in the course of study were high-quality and commercially obtainable.
3
Epithelial-Mesenchymal Transition Assay
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Ham’s F12 nutrient mix, Roswell Park Memorial Institute (RPMI) 1640, fetal bovine serum (FBS), penicillin/streptomycin, and Lab-Tek II 8-well chamber slide were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Corning® Matrigel® Growth Factor Reduced (GFR) basement membrane matrix and cell recovery solution were acquired from Corning (Corning, NY, USA). Ribonuclease A and phenylmethylsulphonyl fluoride (PMSF) protease inhibitor were purchased from Sigma Chemicals (St. Louis, MO, USA). Bradford solution was purchased from Bio-Rad (Hercules, CA, USA). Hoechst 33342, TRITC-conjugated phalloidin, primary antibodies against galectin-3, ZEB-1, N-cadherin, E-cadherin, β-catenin, vimentin, horseradish peroxidase (HRP)-conjugated secondary antibodies against anti-rabbit and anti-mouse antibodies, Alexa Fluor® 647-conjugated secondary antibody against anti-rabbit antibodies and SignalFire™ ECL Reagent were purchased from Cell Signaling Technologies (Denver, MA, USA). The primary antibody against β-actin was purchased from Sigma-Aldrich. Triton X-100 was obtained from Bio-Rad. Recombinant human galectin-3 (rGal-3) was purchased from Merck (Dorset, UK).
4
Western Blot Analysis of HIF-1α
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RIPA buffer (Thermo Fisher Scientific Inc.) containing both protease inhibitor cocktail (Sigma-Aldrich, St Louis, MO, USA) and the PhosSTOP phosphatase inhibitor cocktail (Roche, Tokyo, Japan) was used to lyse cells. Protein concentrations were measured with the BCA kit (Thermo Fisher Scientific Inc.), and equal amounts of extracted proteins were separated on 10% SDS-PAGE gels and transferred onto PVDF membranes (Bio-Rad Inc., Hercules, CA, USA). The membranes were incubated with the indicated primary antibody, followed by the appropriate HRP-conjugated secondary antibody. The bands were detected by chemiluminescence (Thermo Fisher Scientific Inc.). Primary antibody against HIF-1α (diluted 1:1000; HPA001275) was purchased from Sigma-Aldrich, and primary antibody against β-actin was obtained from Sigma Chemical (St Louis, MO, USA).
5
Evaluating K562 Cell Line Responses
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K562 and K562/G01 cell line purchased from Chinese Academy of Medical Sciences (Tianjin, China), were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY) and penicillin-streptomycin in an incubator maintained at 37°C in an atmosphere containing 5% CO2. NC was purchased from Tauto Biotech (Shanghai, China). Primary antibodies for c-Myc and p21 were obtained from Cell Signaling Technology (Beverly, MA). Primary antibody against c-Myc pThr58 was purchased from ImmunoWay (Newark, DE). Primary antibody against globin γ was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Primary antibody against β-actin was from Sigma-Aldrich (St Louis, MO). FITC-conjugated CD71 and PE-conjugated CD235a were from eBioscience (San Diego, CA). All secondary antibodies were obtained from LI-COR Biosciences (Lincoln, NE). Benzidine, GSK3β inhibitors LiCl or Bio, and all other chemicals were obtained from Sigma-Aldrich (St Louis, MO).
6
Epithelial-Mesenchymal Transition Protein Analysis
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The compounds emodin and 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Human recombinant TGF-β was purchased from R&D Systems (Minneapolis, MN, USA). Primary antibodies against Snail, E-cadherin, β-cadherin, MMP-2 and Slug were purchased from Cell Signaling Technology (Beverly, MA, USA). Primary antibody against vimentin was purchased from Abcam Inc. (Cambridge, MA, USA). Primary antibodies against Twist were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Primary antibody against β-actin was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Secondary antibodies, HRP-conjugated Goat anti-Mouse IgG and Goat anti-Rabbit IgG, were obtained from Millipore (Billerica, MA, USA).
7
Cinobufagin-Induced Apoptosis Pathway
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Cinobufagin was purchased from Anhui Jinchan Biochemical Co., Ltd. (Anhui, China). Cell counting kit-8 (CCK-8) was purchased from Dojindo (Kumamoto, Japan). The primary antibody against β-actin was purchased from Sigma (St. Louis, MO, United States). Primary antibodies against caspase-3, caspase-9, Bcl-2, and PARP were purchased from Abcam (Cambridge, United Kingdom).
8
Quercetin-Mediated Apoptosis Induction
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Quercetin, dexamethasone, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI), and ribonuclease A were all purchased from Sigma(St. Louis, MO, USA) Z-VAD-FMK was purchased from R&D systems (USA). Annexin V Apoptosis Detection Kit FITC/PI was obtained from eBioscience (CA, USA). Primary antibodies against caspase-3, caspase-8, caspase-9, poly(ADP-ribose)polymerase 1 (PARP-1), and c-myc were procured from Cell Signaling Technology (MA, USA). Primary antibody against p21 was obtained from Abcam (Cambridge, UK). Primary antibody against β-actin was obtained from Sigma-Aldrich. Horseradish peroxidase (HRP)–conjugated antirabbit and antimouse antibodies were procured from Jackson Immuno Research Laboratories (PA, USA).
9
Antioxidant and DNA Damage Assessment
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EGCG (>99% purity) was obtained from Yibeijia Tea Technology (Hangzhou, China). Melatonin (>98% purity), nitroblue tetrazolium (NBT) and protease inhibitor cocktail were purchased from Sigma (St. Louis, MO, USA). The primary antibody against β-actin was purchased from Sigma (St. Louis, MO, USA). The primary antibody against γ-H2AX was purchased from Santa Cruz (Dallas, TX, USA). The primary antibody against TrxR1 was offered by Professor Gary F. Merrill (Oregon State University). The other primary antibodies as well as the anti-mouse and anti-rabbit secondary antibodies were purchased from Cell Signaling Technology (Boston, MA, USA). L-glutamine was purchased from Gibco (Grand Island, NY, USA). Fetal bovine serum, RPMI 1640 medium, trypsin, penicillin-streptomycin solutions were purchased from HyClone (Logan, UT, USA). 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) was a product of Amresco LLC (Solon, OH, USA). Other chemicals were of the highest grade available.
10
Molecular Signaling Pathway Antibody Panel
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