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Prestained protein markers

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Prestained protein markers are laboratory tools used to estimate the molecular weights of proteins in a given sample. They contain a mixture of pre-stained proteins with known molecular weights, which can be used as reference standards during protein analysis techniques such as gel electrophoresis.

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Lab products found in correlation

Polyvinylidene difluoride pvdf membrane, by Bio-Rad (4 mentions) Enhanced chemiluminescence ecl detection system, by Bio-Rad (3 mentions) Ribozol rna extraction reagent, by Avantor (1 mentions) Luciferase assay system e1501, by Promega (1 mentions) Salmon calcitonin, by Merck Group (1 mentions) β actin monoclonal antibody, by Merck Group (1 mentions) Phosstop, by Roche (1 mentions) Tissue culture dishes and plates, by Corning (1 mentions) Assay reagents and standards, by Bio-Rad (1 mentions) Whatman, by Cytiva (1 mentions) Protran, by Cytiva (1 mentions) Precast gel, by Bio-Rad (1 mentions) Coomassie brilliant blue r 250, by Bio-Rad (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions) Amersham imager 680, by Cytiva (1 mentions) Clarity max, by Bio-Rad (1 mentions) Amersham imager, by Cytiva (1 mentions) Integrin β4, by Santa Cruz Biotechnology (1 mentions) Dc protein assay kit, by Bio-Rad (1 mentions) Cyp24a1, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Protein marker (19 citations) Prestained markers (5 citations)

11 protocols using prestained protein markers

1

Molecular Mechanism of Ellagic Acid Inhibition

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The small molecule inhibitor, ellagic acid dihydrate (TBBD), CARM1 polyclonal antibody (09-818), anti-actin polyclonal antibody (A2066), and salmon calcitonin (05232401) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Sepharose Protein A was obtained from Rockland Immunochemicals, Inc. (Gilbertsville, PA, USA). VDR (C-20) and OPN (P-18) antibodies and anti-rabbit (sc-2004) and anti-goat (sc-2020) horseradish peroxidase-(HRP) conjugated) antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Asymmetric dimethyl H3R17 antibody (ab8284) was obtained from Abcam (Cambridge, MA, USA). Donkey anti-rabbit IgG HRP (711-035-152) was purchased from Jackson ImmunoResearch Laboratories, Inc. (West Grove, PA, USA). 1,25(OH)2D3 was purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). Pre-stained protein markers and polyvinylidene difluoride (PVDF) membranes were obtained from Bio-Rad. The luciferase assay system (E1501) was purchased from Promega (Madison, WI, USA). RiboZol RNA Extraction Reagent was purchased from Amresco (Solon, OH, USA).
Mady L.J., Zhong Y., Dhawan P, & Christakos S. (2023). Role of Coactivator Associated Arginine Methyltransferase 1 (CARM1) in the Regulation of the Biological Function of 1,25-Dihydroxyvitamin D3. Cells, 12(10), 1407.
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2

Comprehensive List of Reagents Used

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The reagents used in this study were purchased from the following suppliers: Prestained protein markers and Bio-Rad assay reagents and standards (Bio-Rad Laboratories, Hercules, CA); Protran BA83 Nitrocellulose membrane (Whatman Inc, Sanford, ME); Pierce ECL (Pierce Biotechnology, Rockford, IL); protease inhibitor cocktail tablets complete EDTA-free and PhosStop (Roche Applied Biosciences, Indianapolis, IN); antibiotic-antimycotic, and trypsin–EDTA (Invitrogen Life Technologies Inc, Carlsbad, CA); Blue X-Ray film (Phenix Research Products, Hayward, CA); fetal bovine serum (HyClone, Logan, UT); and tissue culture dishes and plates (Corning Inc, Corning, NY). Antagonists/inhibitors for EP2 (AH6809), EP4 (AH23848) were purchased from Sigma (Sigma-Aldrich, St-Louis, MO). All other antibodies used in this study were purchased from Cell Signaling Technology (Danvers, MA), Chemicon International (Billerica, MA), or Santa Cruz Biotechnology (Santa Cruz, CA) except β-actin monoclonal antibody (Sigma-Aldrich), goat anti-rabbit or anti-mouse IgG conjugated with horseradish peroxidase (Kirkegaard & Perry Laboratories, Gaithersburg, MA). The chemicals used were molecular biological grade from Fisher Scientific (Pittsburgh, PA) or Sigma-Aldrich (St. Louis, MO).
Arosh J.A., Lee J., Starzinski-Powitz A, & Banu S.K. (2015). Selective Inhibition of Prostaglandin E2 Receptors EP2 and EP4 Modulates DNA Methylation and Histone Modification Machinery Proteins in Human Endometriotic Cells. Molecular and cellular endocrinology, 409, 51-58.
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3

Colorimetric Assay for Azoreductase Activity

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Azoreductase activity assays were performed as described previously [6] (link). The activities of AzoA wild type and mutants were analyzed by measuring the reduction of methyl red at 430 nm at room temperature. The reaction mixture was 1956 μl potassium phosphate buffer (pH 7.1), 2 μl methyl red (25 μM), 20 μl NADH (0.1 mM), 2 μl FMN (0.5 μM), and 20 μl azoreductase. The amount of azoreductase needed for the reduction of 1 μmol methyl red per minute was defined as one unit of activity. Specific azoreductase activity is defined as units of activity per mg protein. The protein was quantified using the Bicinchoninic acid assay (Pierce) with bovine serum albumin (BSA) as the standard. SDS-PAGE was carried out using precast gels (12%, Bio-Rad). Prestained protein markers (Bio-Rad) were used and the gel was stained by Coomassie brilliant blue R-250 (Bio-Rad).
Sun J., Kweon O., Jin J., He G.X., Li X., Cerniglia C.E, & Chen H. (2017). Mutation network-based understanding of pleiotropic and epistatic mutational behavior of Enterococcus faecalis FMN-dependent azoreductase. Biochemistry and Biophysics Reports, 12, 240-244.
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4

Vitamin D Metabolites and Protein Expression

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1,25-Vit D3 and 24,25-Vit D3 were purchased from Enzo Life Sciences (Catalog# BML-DM200-0050, BML-DM300-0050, Farmingdale, NY). Antibodies for CYP24A1 and CYP27B1 were purchased from ABCAM (Catalog# ab203308, ab206655, Cambridge, MA). VDR and Thy1/CD90 antibodies were purchased from Cell Signaling Technology (Catalog# 12550S and 13801S, Danvers, MA). MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) and keratocan antibodies were purchased from ThermoFisher Scientific (Catalog# M6494, and PA5-79552, Waltham, MA). The Pdia3 (Protein disulfide isomerase family A member 3) antibody (Ab099) was a kind gift of Dr. Ilka Nemere. Pdia3 inhibitor LOC14 was purchased from Tocris, Bio-Techne Corporation (Catalog# 5606, Minneapolis, MN). Pre-stained protein markers were obtained from Bio-Rad (Catalog# LC5699, Hercules, CA). Polyvinylidene difluoride (PVDF) membrane and the enhanced chemiluminescence (ECL) detection system were obtained from Bio-Rad (Catalog# 1704272, 1705060, Hercules, CA).
Lu X., Chen Z, & Watsky M.A. (2021). Effects of 1,25 and 24,25 Vitamin D on Corneal Fibroblast Vitamin D Receptor and Vitamin D Metabolizing and Catabolizing Enzymes. Current eye research, 46(9), 1271-1282.
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5

Protein Denaturation and Western Blot

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Proteins were denaturated for 30 min at 65 °C and resolved by SDS-PAGE. Afterward, we performed electrotransfer onto a nitrocellulose membrane. Immunocomplexes were detected using an enhanced chemiluminescence detection system (Clarity or Clarity Max, Bio-rad) on Amersham Imager 680 and 800 as 16-bit grayscale TIFF files. The molecular weight of proteins was estimated with pre-stained protein markers (Bio-rad).
Niedziółka S.M., Datta S., Uśpieński T., Baran B., Skarżyńska W., Humke E.W., Rohatgi R, & Niewiadomski P. (2024). The exocyst complex and intracellular vesicles mediate soluble protein trafficking to the primary cilium. Communications Biology, 7, 213.
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6

Quantitative Analysis of Skin Protein Expression

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1,25(OH)2D3 and 24R,25(OH)2D3 were purchased from Enzo Life Sciences (Farmingdale, NY, USA). Antibodies for DSG1, DSC2, and integrin β4 were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Plectin and integrin α6 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Prestained protein markers were obtained from Bio-Rad (Hercules, CA, USA). Polyvinylidene difluoride (PVDF) membrane and the enhanced chemiluminescence (ECL) detection system were obtained from Bio-Rad.
Lu X, & Watsky M.A. (2019). Influence of Vitamin D on Corneal Epithelial Cell Desmosomes and Hemidesmosomes. Investigative Ophthalmology & Visual Science, 60(13), 4074-4083.
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7

Protein Quantification and Immunoblotting

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Protein determinations were performed using the Bio-Rad D-C protein assay kit with bovine serum albumin as the protein standard. Western blots were carried out as described previously (37 (link)). The molecular sizes of the immunodetected proteins were verified by comparison to the migration of prestained protein markers (Bio-Rad) electrophoresed in parallel lanes (39 (link)). Antibodies to PAF53 and mammalian A127 have been described previously (37 (link)). The antibodies to GFP were obtained from Clontech, the antibodies to GST were obtained from Sigma.
Penrod Y., Rothblum K., Cavanaugh A, & Rothblum L.I. (2014). Regulation of the Association of the PAF53/PAF49 Heterodimer with RNA Polymerase I. Gene, 556(1), 61-67.
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8

Vitamin D Metabolite Analysis Techniques

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1,25(OH)2D3 and 24R,25(OH)2D3 were purchased from Sigma (Ann Arbor, MI). CYP24A1 and CYP27B1 polyclonal antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). VDR and GAPDH antibodies were purchased from Cell Signaling Technology (Danvers, MA). Prestained protein markers were obtained from Bio-Rad (Hercules, CA). Polyvinylidene difluoride (PVDF) membrane and the enhanced chemiluminescence (ECL) detection system were obtained from Bio-Rad (Hercules, California).
Lu X., Chen Z., Mylarapu N, & Watsky M.A. (2017). Effects of 1,25 and 24,25 Vitamin D on Corneal Epithelial Proliferation, Migration and Vitamin D Metabolizing and Catabolizing Enzymes. Scientific Reports, 7, 16951.
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9

Caspase-3 and Caspase-8 Activity Assay

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Smac mimetic LCL161, recombinant (active) human caspase-3 (ALX-201-059), substrates Ac-DEVD-MCA (caspase-3), and Ac-IETD-MCA (caspase-8), and inhibitor (zVAD-fmk) were from Sapphire Bioscience (Sapphire Bioscience Pty Ltd). Recombinant (active) caspase-8 was expressed and purified from E. coli using established procedures (41 (link)). Anti-phospho-MLKL (ab196436) antibody was from Abcam, and anti-caspase-8 (1C12, #9746) and anti-cleaved caspase-8 (small subunit, #9748) were from Cell Signaling Technology. Nitrocellulose membrane and ECL Select Western Blotting Detection Reagent were from GE Healthcare. Secondary antibodies were from DAKO (Agilent). All cell culture media: Dulbecco modified Eagle medium, Roswell Park Memorial Institute 1640 (RPMI) medium, Medium 199 (M199), fetal bovine serum (FBS), penicillin/streptomycin (Pen/Strep) and TrypLE Express, and human TNF-α were from Thermo Fisher Scientific. Precast Mini-Protean gels (including TGX Stain-Free) and prestained protein markers were from Bio-Rad Laboratories Inc. All other reagents, including anti-MLKL (MABC604) and anti-β-actin (A5316) antibodies, were from Merck.
Bozonet S.M., Magon N.J., Schwartfeger A.J., Konigstorfer A., Heath S.G., Vissers M.C., Morris V.K., Göbl C., Murphy J.M., Salvesen G.S, & Hampton M.B. (2023). Oxidation of caspase-8 by hypothiocyanous acid enables TNF-mediated necroptosis. The Journal of Biological Chemistry, 299(6), 104792.
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10

Sorghum Grain Protein Extraction

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Whole food-grade retail sorghum grain (Sorghum bicolor Moench) was purchased from Northern
Food Inc. (Hicksville, NY, USA). Commercial zein (260-01283) was purchased
from Wako Pure Chemical Industries, Ltd. (New York, NY, USA). Tris-alkali,
sodium dodecyl sulfate (SDS), N,N,N′,N′-tetramethylethylenediamine
(TEMED), 2-mercaptoethanol (2-ME), bromophenol blue, Coomassie brilliant
blue G-250, and prestained protein markers (broad range protein maker)
were purchased from Bio-Rad Laboratories, Inc. (Hercules, CA, USA).
Milli-Q water was used throughout the experiments.
Xiao J., Li Y., Li J., Gonzalez A.P., Xia Q, & Huang Q. (2014). Structure, Morphology, and Assembly Behavior of Kafirin. Journal of Agricultural and Food Chemistry, 63(1), 216-224.
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