HEp-2 and A549 cells were purchased from the ATCC (Manassas, VA), and were maintained in F12K and MEM medium respectively, containing 10% (v/v) FBS. SAECs, isolated from the normal human lung distal portion, were purchased from Lonza (Pittsburgh, PA), and were maintained according to the manufacturer’s protocol. RSV long strain was propagated in HEp-2 cells at 37 °C and purified by sucrose gradient as previously described.58 (link), 70 Viral titer was determined by immunostaining in HEp-2 cells using polyclonal biotin-conjugated goat anti-RSV antibody (7950–0104; Bio-Rad, Hercules, CA) and streptavidin peroxidase polymer (S2438; Sigma-Aldrich, St. Louis, MO) sequentially, as previously described.58 (link), 70 The polyclonal biotin-conjugated goat anti-RSV antibody was also used for Western blot to detect viral protein expression. The monoclonal antibody against β-actin was from Sigma (A1978). Primary antibodies against IRF-3(CST#4302), phospho-IRF3(Ser396)(#29047), p65(CST#4764), phospho-p65(Ser276)(CST#3037), and phospho-p65(Ser536)(CST# 3033) were purchased from Cell Signaling Technology (Denvers, MA), and goat anti-rabbit IgG-HRP (4050–05) was purchased from SouthernBiotech (Birmingham, AL).
Monoclonal antibody against β actin
Manufactured by Merck Group
Sourced in United States
Monoclonal antibody against β-actin is a laboratory reagent used to detect and quantify the presence of the β-actin protein in cell and tissue samples. β-actin is a ubiquitous cytoskeletal protein found in all eukaryotic cells and is often used as a reference or control in various experimental techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Liquid dab substrate chromogen system horseradish peroxidase, by Agilent Technologies (2 mentions)
Sodium chloride (nacl), by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (2 mentions)
Glycine, by Merck Group (2 mentions)
Revertaid first strand cdna synthesis kit, by Vazyme (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Phospho irf3 ser396, by Cell Signaling Technology (1 mentions)
Phospho p65 ser536, by Cell Signaling Technology (1 mentions)
Goat anti rabbit igg hrp, by Southern Biotech (1 mentions)
Streptavidin peroxidase polymer, by Merck Group (1 mentions)
Saecs, by Lonza (1 mentions)
Antibody against ho 1, by Abcam (1 mentions)
Muc5ac antibody, by Thermo Fisher Scientific (1 mentions)
Sp600125, by Enzo Life Sciences (1 mentions)
Phospho egfr tyr1068, by Cell Signaling Technology (1 mentions)
Ici 182 780, by Merck Group (1 mentions)
26 protocols using monoclonal antibody against β actin
1
Respiratory Syncytial Virus Propagation and Detection
2
Antibody Characterization for Signaling Pathways
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SP600125 was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). Antibodies against phospho-ERK 1/2 (Thr-202/Tyr204), phospho-p38 MAPK (Thr-180/Tyr-182), phospho-JNK 1/2 (Thr183/Tyr185) and phospho-EGFR (Tyr1068) were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Monoclonal antibody against β-actin was purchased from Sigma-Aldrich Japan (Tokyo, Japan). Antibody against γ-tubulin was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). MUC5AC antibody was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Antibody against HO-1 was purchased from Abcam (UK).
3
Immunocytochemical Analysis of Molecular Markers
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ICI 182,780 and E2 were obtained from Sigma (Madison, WI). Polyclonal antibodies against F4/80 and integrin-α5β1 were obtained from Abcam. Anti-BrdU monoclonol antibody came from BD Biosciences (San Jose, CA). The liquid DAB+ substrate chromogen system-horseradish peroxidase used for immunocytochemistry was obtained from Dako Cytomation (Carpinteria, CA). Penicillin, streptomycin and fetal bovine serum (FBS) were obtained from Invitrogen (Carlsbad, CA). Tris, glycine, NaCl, SDS, bovine serum albumin, and monoclonal antibody against β-actin were obtained from Sigma (Madison, WI).
4
Autophagy and Apoptosis Regulation
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The monoclonal anti-Beclin-1 antibody and high-mobility group box1(HMGB1) were purchased from Santa Cruz. The polyclonal rabbit anti-microtubule-associated protein 1A/1B-light chain 3 (LC3) and a monoclonal antibody against β-Actin were obtained from Sigma. The following antibodies were purchased from Cell Signaling Technology: cleaved caspase-3 and phospho-specific antibodies to mTOR (Ser2448), Akt (Ser473) and p70 ribosomal protein S6 kinase (S6K) (Thr389), along with total antibodies directed against mTOR, Akt, and p70S6K. Rapamycin and chloroquine (CQ) were purchased from Sigma.
5
Immunoblotting of TREM-1 and M-CSF in OIR
6
Western Blot Analysis of Aminoacyl-tRNA Synthetases
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Spinal dorsal horn (L4–5) tissue samples were homogenized in modified radioimmunoprecipitation assay buffer [RIPA; 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 0.5% deoxycholic acid, 0.5% Triton X-100, 1 mM phenylmethylsulfonyl fluoride, 1 mM sodium o-vanadate, 1× protease inhibitor cocktail tablet (Roche Molecular Biochemicals, Nutley, NJ, USA)]. After centrifugation for 30 minutes at 4°C, the supernatant was used for western blot analysis.
Western blot analysis was performed according to a previous study (Seo et al., 2014). We used the following primary antibodies: a polyclonal rabbit anti-FARSB (1:200; Proteintech, Chicago, IL, USA), a polyclonal rabbit anti-IARS (1:200; Neomics, Seoul, Korea), a polyclonal rabbit anti-MARS (Neomics, 1:200), a polyclonal rabbit anti-TARS (1:200; Gentex, San Antonio, TX, USA) and a monoclonal antibody against β-actin (1:5,000; Sigma). For quantification, images were scanned and analyzed with the LAS image analysis system (Fujifilm, Tokyo, Japan). Optical density (band intensity) ratios of FARSB, IARS, MARS and TARS to β-actin were calculated.
Western blot analysis was performed according to a previous study (Seo et al., 2014). We used the following primary antibodies: a polyclonal rabbit anti-FARSB (1:200; Proteintech, Chicago, IL, USA), a polyclonal rabbit anti-IARS (1:200; Neomics, Seoul, Korea), a polyclonal rabbit anti-MARS (Neomics, 1:200), a polyclonal rabbit anti-TARS (1:200; Gentex, San Antonio, TX, USA) and a monoclonal antibody against β-actin (1:5,000; Sigma). For quantification, images were scanned and analyzed with the LAS image analysis system (Fujifilm, Tokyo, Japan). Optical density (band intensity) ratios of FARSB, IARS, MARS and TARS to β-actin were calculated.
7
Corneal Epithelial Cell Culture Protocol
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Human corneal epithelial cells were obtained from L.V. Prasad Eye Institute, Hyderabad India. T-25, T-75 flasks and culture dishes were purchased from Corning Life Sciences, USA. Fetal bovine serum (FBS) and Rhodamine 123 were procured from GIBCO-BRL Life Technologies, USA. Antibodies to Cyt c, Bcl-2, Bax, NF-κB, I-κB, p27 were purchased from Upstate Biotechnology, USA. COX-2 antibody was obtained from Cayman Chemicals, USA. COX-2 siRNA was from Santacruz Biotechnology, USA. Universal negative control (NC1) siRNA was from IDT, USA. Western blotting detection reagent was purchased from GE Healthcare Life Sciences, USA. cDNA synthesis kit and Lipofectamine 2000 was ordered from Invitrogen, USA. MEM alpha, EGF, Insulin, nutrient mixture F12, Pseudomonas aeruginosa LPS, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide], BCIP/NBT, TMB/H2O2, TRIZOL reagent, monoclonal antibody against β-actin were purchased from Sigma, USA. All other chemicals and reagents of molecular biology grade were purchased from local companies in India.
8
Quantitative Western Blot Analysis of AQP4
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The isolated hemisphere was homogenized in a mixture of 250 mmol/L sucrose, 10 mmol/L Tris-HCl, pH 7.4, 0.2 mmol/L EDTA and 20 g/ml phenylmethyl-sulfonyl fluoride. Ten µg of soluble cortical and striatal proteins were resolved using 12.5% self-cast SDS-polyacryamide gel (SDS-PAGE) electrophoresis and subsequently transferred onto polyvinylidene difluoride (PVDF) membranes (Bio-Rad) m. The blot was then incubated overnight at 4°C with either a polyclonal antibody against AQP4 (1∶1000, Cell Signal Technology) or a monoclonal antibody against β-actin (1∶25,000, Sigma) in TBST (10 mmol/L Tris, pH 8.0,150 mmol/L NaCl, 0. 05% Tween 20) containing 3% skimmed milk. After washing, the membrane was incubated with horseradish peroxidase labeled antibody (1∶1000, Sigma) for 1 hour at room temperature. The membrane was incubated in ECL solution, and the gel image was captured using an imaging camera Bio-Rad and analyzed using gel image system (Quantity one) to estimate the integral optical density of the protein bands.
9
Investigating TGF-β Signaling in Human Cancer Cells
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The human HCC cell lines HepG2, Hep3B and Human Umbilical Vein Endothelial Cells (HUVEC) were obtained and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco BRL, supplemented with 10% foetal bovine serum, 100 mg/ml penicillin, and 100 mg/ml streptomycin. The cells were incubated at 37°C in a humidified atmosphere at 5% CO2. The TGF-b receptor kinase inhibitor LY2109761 was purchased from Eli Lilly (Indianapolis, IN). Cell Counting kit-8 was purchased from Sigma (Milan, Italy), monoclonal blocking antibody against human E-cadherin, SHE78–7, from Alexis (Lausanne, Switzerland) and polyclonal antibodies against phospho-Smads were purchased from Cell Signaling Technology Inc. (Danvers, MA) and monoclonal antibody against β-actin were purchased from Sigma, while growth factor–reduced (GFR) Matrigel were purchased from BD.
10
Antibody Characterization and Detection
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The monoclonal antibody against β-actin was provided by Sigma-Aldrich (St. Louis, MO). Anti-HA monoclonal and polyclonal, anti-myc monoclonal and anti-DDDDK tag (FLAG tag) antibodies were provided by MBL (Nagoya, Japan). Anti-mouse IgG Alexa 546 and anti-rabbit IgG Alexa 488 conjugated antibodies were provided by Thermo Fischer Scientific (Waltham, MA)
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