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Acrylamide bis solution 29 1

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30% Acrylamide/Bis Solution 29:1 is a laboratory reagent used in the preparation of polyacrylamide gels for electrophoresis. This solution contains a 29:1 ratio of acrylamide to bis-acrylamide at a concentration of 30% w/v.

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Lab products found in correlation

Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions) Imagequant 800 system, by Cytiva (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Protease inhibitor, by Roche (1 mentions) Ammonium persulfate, by Thermo Fisher Scientific (1 mentions) Trypsin from bovine pancreas, by Merck Group (1 mentions) Glacial acetic acid, by Thermo Fisher Scientific (1 mentions) Lc ms grade water, by Merck Group (1 mentions) Tris hcl, by Bio-Rad (1 mentions) Bio safe coomassie brilliant blue g 250 stain, by Bio-Rad (1 mentions) Salivary alpha amylase kinetic enzyme assay kit, by Salimetrics (1 mentions) Hplc grade acetonitrile, by Thermo Fisher Scientific (1 mentions) Iodoacetamide, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Ammonium bicarbonate, by Merck Group (1 mentions) Laemmli sample buffer, by Bio-Rad (1 mentions) Bicinchoninic acid protein assay kit, by Merck Group (1 mentions) Coomassie brilliant blue g 250, by Bio-Rad (1 mentions) Ace from porcine kidney, by Merck Group (1 mentions) Precision plus protein dual color standard, by Bio-Rad (1 mentions)

Close spelling variants of this product

Acrylamide–bis-acrylamide solution (29∶1; Acrylamide:bis 29:1 Acrylamide/bis solution Acrylamide/bis-acrylamide solution Bis-acrylamide Acrylamide solution Acrylamide

4 protocols using acrylamide bis solution 29 1

1

Western Blot Analysis of Inflammatory Signaling

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Isolated Mo cultured at 500,000 cells/well in the presence of different stimuli were lysed using RIPA buffer 1% of phosphatase and protease inhibitors (Roche Diagnostics) with 10 min of sonication. Protein lysates were resolved in a 10% acrylamide (30% Acrylamide/Bis Solution 29:1, Bio Rad) gel with SDS and transferred to a nitrocellulose membrane (Fisher Scientific). Afterwards, membranes were blocked with 4% BSA (Sigma-Aldrich) in Tris-Buffered saline and incubated at 4 °C overnight with primary antibodies directed either to phospho-p65 (Cell Signaling ref 3039), anti-p65 (ab32536, E379, Abcam;), anti-caspase-1 (AF6215; R&D Systems), anti-Gasdermin D (69469, E5O4N; Cell Signaling), anti-IL-1β (12242, 3A6; Cell Signaling) or anti-GAPDH (FF26A/F9; BioLegend) following manufacturer’s specifications. Each primary antibody was used at a 1:1000 dilution. Subsequently, the membranes were washed and incubated at RT for 1 h with the respective anti-rabbit (31460; Thermofisher; dilution 1:2000), anti-goat (81–1620; Thermofisher; dilution 1:2000) or anti-mouse (31430; Thermofisher; dilution 1:5000) secondary antibodies. Chemiluminescence of protein band intensity was quantified by ImageQuant 800 system (Amersham) using the IQ800 V1.2.0 Software.
Tsukalov I., Sánchez-Cerrillo I., Rajas O., Avalos E., Iturricastillo G., Esparcia L., Buzón M.J., Genescà M., Scagnetti C., Popova O., Martin-Cófreces N., Calvet-Mirabent M., Marcos-Jimenez A., Martínez-Fleta P., Delgado-Arévalo C., de los Santos I., Muñoz-Calleja C., Calzada M.J., González Álvaro I., Palacios-Calvo J., Alfranca A., Ancochea J., Sánchez-Madrid F, & Martin-Gayo E. (2024). NFκB and NLRP3/NLRC4 inflammasomes regulate differentiation, activation and functional properties of monocytes in response to distinct SARS-CoV-2 proteins. Nature Communications, 15, 2100.
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2

EBNA1 DNA-binding domain analysis

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Purified untagged EBNA1 WT DNA-binding domain (459–607) or its point mutant derivatives were incubated at the indicated concentrations (6.25 nM to 200 nM) with DNA probes (160 femtomoles). Oligonucleotide DNA probes were end-labeled with 32P-γ-ATP and T4 polynucleotide kinase (PNK), and annealed in annealing buffer (50mM Tris HCl (pH 7.5); 400mM NaCl) as described (Rass and West, 2006 (link)). Oligonucleotides were purchased from IDT Inc. in standard desalted form. A list of oligonucleotides in provided in Key Resources Table. Binding reactions were incubated for 10 mins at room temperature in 20 μl binding buffer containing 10 mM HEPES pH 7.5, 5 mM MgCl2, 2.5 mM DTT, 0.25% Tween, 200 mM NaCl, and 10% glycerol. DNA-protein complexes were resolved by electrophoresis at 250 Volts in 5% non-denaturing polyacrylamide gels (Bio-rad, 30% Acrylamide/Bis Solution 29:1, 1610156) in 0.5X Tris borate-EDTA buffer. The gels were subjected to autoradiography on X-ray films followed by exposure to phosphorimager screens, scanned in Typhoon, and signals quantified using ImageQuant software.
Dheekollu J., Wiedmer A., Ayyanathan K., Deakyne J.S., Messick T.E, & Lieberman P.M. (2021). Cell Cycle-Dependent EBNA1-DNA Cross-Linking Promotes Replication Termination at oriP and Viral Episome Maintenance. Cell, 184(3), 643-654.e13.
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3

Histatin-3 Quantification in Saliva

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Ammonium persulfate, glacial acetic acid and HPLC-grade acetonitrile were obtained from Fisher Scientific (Pittsburgh, PA). Ammonium bicarbonate, β-mercaptoethanol, trypsin from bovine pancreas, trifluoroacetic acid (TFA), formic acid (FA), dithiothreitol, iodoacetamide, and LC-MS-grade water were purchased from Sigma (St. Louis, MO). Bio-Safe Coomassie Brilliant Blue G-250 stain and 2x Laemmli sample buffer were from Bio-Rad (Hercules, CA). For polyacrylamide gel casting, 30% acrylamide/bis solution (29:1), 1.5 M Tris-HCl (pH 8.8), 0.5 M Tris-HCl (pH 6.8), and N,N,N’,N’-tetramethylelthylenediamine (TEMED) were also obtained from Bio-Rad. Monoclonal histatin-3 antibody (4G9) was purchased from Novus Biologicals (Littleton, CO). Histatin antibody (H-40), rabbit polyclonal IgG, was obtained from Santa Cruz Biotechnology. Synthetic histatin-3 was from Genemed Synthesis Inc. (San Antonio, TX). A salivary alpha-amylase kinetic enzyme assay kit was purchased from Salimetrics (State College, PA).
Marvin R.K., Saepoo M.B., Ye S., White D.B., Liu R., Hensley K., Rega P., Kazan V., Giovannucci D.R, & Isailovic D. (2017). Salivary protein changes in response to acute stress in medical residents performing advanced clinical simulations: a pilot proteomics study. Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals, 22(3-4), 372-382.
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4

Casein Purification and Characterization

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Casein (Promilk 85) was obtained from Ingredia (Arrax Cedex, France). ACE from porcine kidney, bicinchoninic acid protein assay kit, Hammarsten casein and Triton X-100 were purchased from Sigma (St. Louis, MO, USA).
Glucose was obtained from Panreac (Barcelona, Spain), bacteriological peptone was purchased from Cultimed (Barcelona, Spain) and yeast extract and agar were acquired from Pronadisa (Madrid, Spain). The fluorogenic substrate o-aminobenzoyl-Gly-p-nitro-Phe-Pro was provided by Bachem Feinchemikalien (Bubendorf, Switzerland). Coomassie Brilliant Blue G-250, 30% acrylamide/bis solution 29:1 and Precision Plus Protein Dual Color standards were obtained from Bio-Rad Laboratories (Hercules, CA, USA).
García-Tejedor A., Sánchez-Rivera L., Recio I., Salom J.B, & Manzanares P. (2015). Dairy Debaryomyces hansenii strains produce the antihypertensive casein-derived peptides LHLPLP and HLPLP. Lebensmittel-Wissenschaft + [i.e. und] Technologie. Food science + technology. Science + technologie alimentaire, 61(2).
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