Pageblue protein staining solution

Manufactured by Thermo Fisher Scientific

Sourced in United States, Lithuania, Germany

PageBlue Protein Staining Solution is a ready-to-use liquid stain for the detection of proteins in polyacrylamide gels. It is a simple and effective solution for visualizing protein bands after electrophoresis.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

PageBlue protein staining PageBlue Staining solution PageBlue solution PageBlue

131 protocols using pageblue protein staining solution

Purification and Characterization of Est6

Check if the same lab product or an alternative is used in the 5 most similar protocols

The total cell soluble proteins produced by E. coli BL21 Star (DE3)/pPAL7-Est6 and purified Est6 dialyzed and concentrated either in 50 mM sodium phosphate buffer (pH 7.0) or 50 mM Tris-HCl buffer (pH 8.0) were analyzed by electrophoresis using 4–12% sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE, NuPAGE Novex Bis-Tris precast gel, Invitrogen, Carlsbad, CA, USA) and the MOPS running buffer (NuPAGE MOPS SDS running buffer, Invitrogen, Carlsbad, CA, USA). The protein fractions were visualized with the PageBlue protein staining solution (Fermentas, Vilnius, Lithuania).
Zymographic analysis of the purified Est6 was performed by native PAGE (NativePAGE Novex 3–12% Bis-Tris Gels; NativePAGE Running Buffer, Invitrogen, Carlsbad, CA, USA). After electrophoresis, the gel was laid on an agarose gel prepared with agarose (1.5%) and tributyrin emulsion (1% tributyrin and 0.1% Tween 80) in a 50 mM Tris-HCl buffer (pH 8.0) and was incubated at 4 °C overnight, and the clear bands appeared. The other corresponding gel was visualized with a PageBlue protein staining solution (Fermentas, Vilnius, Lithuania).

Liaw R.B., Chen J.C, & Cheng M.P. (2022). Molecular Cloning and Characterization of a New Family VI Esterase from an Activated Sludge Metagenome. Microorganisms, 10(12), 2403.

+ Open protocol

+ Expand

SDS-PAGE and BN-PAGE Protein Separation

Check if the same lab product or an alternative is used in the 5 most similar protocols

For SDS-PAGE gel electrophoresis, 15 μl sample was mixed with 5 μl SDS protein gel loading solution (2x) (Quality Biological, MD, USA) and loaded onto a NuPAGE™ 4–12% Bis-Tris gel with 1.5 mm × 10 or 1.5 mm × 15 wells (Novex, ThermoFisher Scientific, NY, USA). The PageRuler™ Plus prestained protein ladder (ThermoFisher Scientific, NY, USA) was used as a molecular marker. The running buffer was NuPAGE MES SDS buffer (20x) (Novex, ThermoFisher Scientific, NY, USA). Gel electrophoresis was performed according to the manufacturer’s instructions. Gels were stained for 1h or overnight with PageBlue™ protein staining solution (ThermoFisher Scientific, NY, USA) and destained for several hours with deionized water.
Assembly and disassembly of GroEL was monitored using blue native PAGE (BN-PAGE; Native PAGE™ 3–12% Bis-Tris gel) with 1 mm × 15 wells. 7.5 μl sample was mixed with 2.5 μl loading solution (NativePAGE 4x sample buffer). The gel, cathode buffer (20x), running buffer (20x), and loading solution were obtained from Novex, ThermoFisher Scientific (NY, USA). Gel electrophoresis was performed in the cold room following the manufacturer’s instructions, followed by staining for 1h or overnight with PageBlue™ protein staining solution (ThermoFisher Scientific, NY, USA) and destaining for several hours with deionized water.

Wälti M.A, & Clore G.M. (2017). Disassembly/Reassembly Strategy for the Production of Highly Pure GroEL, a Tetradecameric Supramolecular Machine, Suitable for Quantitative NMR, EPR and Mutational Studies. Protein expression and purification, 142, 8-15.

+ Open protocol

+ Expand

Affinity Purification and Proteolytic Cleavage of Tagged Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Wild-type His₆-MBP-EYHAGA↓GVVETP-mEYFP substrates attached to Ni-NTA magnetic agarose beads (SAMBs) were suspended in elution buffer (50 mM NaH₂PO₄, 300 mM NaCl, 500 mM imidazole, 0.05% Tween 20, pH 8.0) or cleavage buffer (50 mM NaH₂PO₄, 300 mM NaCl, 0.05% Tween 20, pH 7.5). SAMBs in cleavage buffer were digested by TEV PR (at 7.1 µM final concentration) and VEEV nsP2pro-2 (at 5.2 µM final concentration) on 30 °C overnight. TEV (S219V) PR stock solution was purified according to Kapust et al. [28 (link)].
For native PAGE analysis, samples were prepared from 10 µL of each reaction with 2× loading buffer (62.5 mM Tris-HCI, pH 6.8, 25% glycerol, 0.01% bromophenol Blue). After electrophoresis, the gels were illuminated by using Dark Reader Blue Transilluminator (Labgene Scientific, Châtel-Saint-Denis, Switzerland), and subsequently proteins were detected by PageBlue Protein Staining Solution (Thermo Fisher Scientific, Waltham, MA, USA).
For reducing SDS-PAGE, samples were supplemented with 6x loading buffer (300 mM Tris, pH 6.8, 20% glycerol, 0.05% bromophenol blue, 12% SDS, 100 mM β-mercaptoethanol), denatured at 95 °C for 10 min, followed by electrophoresis. Proteins were detected in the gels by PageBlue Protein Staining Solution (Thermo Fisher Scientific, Waltham, MA, USA).

Bozóki B., Mótyán J.A., Hoffka G., Waugh D.S, & Tőzsér J. (2020). Specificity Studies of the Venezuelan Equine Encephalitis Virus Non-Structural Protein 2 Protease Using Recombinant Fluorescent Substrates. International Journal of Molecular Sciences, 21(20), 7686.

+ Open protocol

+ Expand

In vitro Oxidation and Repair of SurA

Check if the same lab product or an alternative is used in the 5 most similar protocols

SurA-Strep was oxidized in vitro by incubating the purified protein (200 μM) for 3 h at 30°C with 100 mM H₂O₂ in a buffer containing 50 mM NaPi, pH 8.0, 150 mM NaCl. H₂O₂ was then removed by gel filtration using a NAP-5 column (GE healthcare) equilibrated with 50 mM NaPi, pH 8.0, 150 mM NaCl. For the in vitro repair of oxidized SurA (SurA ox), the oxidized protein (30 μM) was incubated with purified MsrP-His₆ (30 μM), 10 mM benzyl viologen and 10 mM of sodium dithionite at 37°C for 1 h. Following repair, SurA was purified by passing the sample through a gravity flow column containing 200 μl Strep-Tactin Sepharose beads (from a 50% suspension, IBA GmbH), previously equilibrated with buffer A (Tris-HCl 100 mM, pH 8.0, NaCl 150 mM, EDTA 1 mM). After washing with buffer A, repaired SurA was eluted using buffer A containing 2.5 mM desthiobiotin. The elution fractions were pooled and submitted to buffer exchange using a NAP-5 column (GE healthcare) equilibrated with 50 mM NaPi, pH 8.0, 150 mM NaCl. To check for the correct oxidation, repair, and purification of SurA, samples were loaded on a SDS-PAGE gel and the proteins visualized with the PageBlue^™ Protein Staining Solution (Fermentas).

Gennaris A., Ezraty B., Henry C., Agrebi R., Vergnes A., Oheix E., Bos J., Leverrier P., Espinosa L., Szewczyk J., Vertommen D., Iranzo O., Collet J.F, & Barras F. (2015). Repairing oxidized proteins in the bacterial envelope using respiratory chain electrons. Nature, 528(7582), 409-412.

+ Open protocol

+ Expand

Sodium Channel Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were denatured by boiling in SDS-PAGE buffer and resolved using 4–20% gradient polyacrylamide gels and followed by either Coomassie brilliant blue G-250 staining with PageBlue protein staining solution (Fermentas) or electrophoretically transferred to Hybond P membrane (Amersham Pharmacia) for Western blot. Membranes were probed with rabbit polyclonal anti-NaV antibody conjugated to HRP diluted 1 : 5000. The membranes were developed by chemiluminescence using the ECL plus detection reagent (Amersham Pharmacia).

Mathew L.G., Herbst-Kralovetz M.M, & Mason H.S. (2014). Norovirus Narita 104 Virus-Like Particles Expressed in Nicotiana benthamiana Induce Serum and Mucosal Immune Responses. BioMed Research International, 2014, 807539.

+ Open protocol

+ Expand

SDS-PAGE and Proteomic Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

SDS-PAGE was performed with 4%–20% precast gels in Tris-based buffer system from SERVA Electrophoresis (#43289.01) according to the manufacturer’s instructions. PageBlue Protein Staining Solution (Fermentas, #R0571) was used for coomassie-staining the gels, images were taken with iPhone 6s (Apple) and the contrast was adjusted using ImageJ (version 1.52d). Global identification and quantification of MSU- and zymosan-binding proteins in the presence of different donor sera was done as previously described (25 (link)). In brief, eluted proteins were reduced with DTT, alkylated with acrylamide, and separated using SDS-PAGE (4%–20%, Sigma-Aldrich). Whole lanes were cut into three individual slices and proteins therein were in-gel digested with trypsin. Generated peptides were analyzed using an LC-MS system consisting of an Orbitrap Velos mass spectrometer coupled to an Ultimate 3000 RSLC nanoflow system (Thermo Fisher Scientific). Raw data were analyzed with the Andromeda search engine implemented in MaxQuant software (version 1.5.3.30; www.maxquant.org). Proteins were identified based on a false discovery rate (FDR) of less than 0.01 on protein and peptide level.

Alberts A., Klingberg A., Hoffmeister L., Wessig A.K., Brand K., Pich A, & Neumann K. (2020). Binding of Macrophage Receptor MARCO, LDL, and LDLR to Disease-Associated Crystalline Structures. Frontiers in Immunology, 11, 596103.

+ Open protocol

+ Expand

In vitro Oxidation and Repair of SurA

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Purification of Human HINT1 and HINT2

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human HINT1 protein was expressed in bacteria E. coli BL21* using the pSGA02-hHINT1 plasmid and purified by AMP -agarose (matrix spacer 8 over the C-8 atom of 5′-AMP, SIGMA, Oakville, ON, USA) affinity chromatography as previously described for rabbit Hint1 [15 (link)]. Human HINT2 was expressed in E.coli using the pGAT2-HINT2 vector and purified in two steps as described [21 (link)]. Briefly, the His6-GST-tagged HINT2 obtained in the first step was purified by Ni- IDA agarose chromatography (Qiagen, Germantown, MD, USA), and then after cleavage by thrombin (to remove the His6-GST-tag) by AMP-agarose chromatography. Lysis and purification of both proteins was performed with the addition of protease inhibitor cocktail (Roche Diagnostics, Mannheim, Germany). Finally, the homogeneous enzyme preparations were concentrated and stored at −80 °C. The purity of the proteins was evaluated by SDS-PAGE analysis and subsequent staining with the PAGE Blue Protein Staining Solution (Fermentas, Lithuania).

Krakowiak A., Czernek L., Pichlak M, & Kaczmarek R. (2022). Intracellular HINT1-Assisted Hydrolysis of Nucleoside 5′-O-Selenophosphate Leads to the Release of Hydrogen Selenide That Exhibits Toxic Effects in Human Cervical Cancer Cells. International Journal of Molecular Sciences, 23(2), 607.

+ Open protocol

+ Expand

Protein Extraction and Fractionation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell extracts were prepared by heating cells resuspended in sample buffer at 100°C for 10 min. The cells were fractionated on a 4%–15% gel (BioRad) and the gels were stained using PageBlue protein staining solution (Fermentas).

Ojha S, & Jain C. (2020). Dual-level autoregulation of the E. coli DeaD RNA helicase via mRNA stability and Rho-dependent transcription termination. RNA, 26(9), 1160-1169.

+ Open protocol

+ Expand

E. coli Outer Membrane Protein Fractionation

Check if the same lab product or an alternative is used in the 5 most similar protocols

E. coli proteins were separated into soluble, inner membrane and OM fractions as previously reported [19 (link)]. The resulting outer membrane protein samples were separated using SDS-PAGE on 10 % Bis-Tris gels (NuPage, Invitrogen). The gels were then stained using PageBlue protein staining solution (Fermentas) according to the manufacturer’s protocol.

Gustavsson M., Do T.H., Lüthje P., Tran N.T., Brauner A., Samuelson P., Truong N.H, & Larsson G. (2015). Improved cell surface display of Salmonella enterica serovar Enteritidis antigens in Escherichia coli. Microbial Cell Factories, 14, 47.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!