Nupage protein gel

Manufactured by Thermo Fisher Scientific

Sourced in United States

NuPAGE protein gel is a pre-cast gel system designed for the separation and analysis of proteins. It provides a consistent and reliable platform for protein electrophoresis.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

NuPAGE 4–12% Bis-Tris proteins gels NuPAGE Bis-Tris SDS/PAGE Protein Gels Novex NuPAGE Precast Protein Gels NuPAGE® Novex® Bis-Tris Protein Gels NuPAGE Novex 4–12% bis-tris midi protein gels NuPAGE Novex Bis-Tris Protein gradient polyacrylamide gels NuPAGE™ 4–12% Bis-Tris Midi Protein Gels NuPAGE™ 4 to 12% Bis-Tris Mini Protein Gels Novex NuPAGE 10% Bis-Tris Protein gels Gradient NuPAGE 4–12% Bis-Tris Protein Gels

9 protocols using nupage protein gel

Western Blot Analysis of Protein Signaling

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

Complete RIPA buffer was prepared by adding 1× Protease and Phosphatase inhibitor Cocktail (Thermo Scientific, Dreieich, Germany) with Pierce RIPA buffer (Thermo Scientific). Complete RIPA buffer were directly added to the cell culture plates or snap frozen tissues and then homogenized to isolate protein. The protein was denatured using 4× LDS sample buffer (Thermo Scientific) and 10× Reducing agent (Thermo Scientific). NuPAGE protein gels (Invitrogen, Carlsbad, CA, USA) were used to run the protein and PVDF membrane (Thermo Scientific) was used to transfer the protein from gel. Membranes were incubated with primary antibodies for overnight at 4 °C and with secondary antibodies for 1 h at RT. ECL substrate (GE Healthcare, Chicago, IL, USA) were added directly into the membrane for 2–3 min before capturing image at Amersham Imager 680 (GE Healthcare). Band intensities were also quantified using the software provided with the Imager.
Primary antibodies—Rabbit IRAG1, Rabbit PKG1α, Rabbit PKG1β (kindly provided by Prof. Schlossmann) [15 (link),25 (link)], Rabbit IP₃RI, Rabbit Pan-Actin (Cell Signaling Technology), Rabbit Serca2a, Rabbit pPLN-Thr 17, Mouse PLN (Badrilla, Leeds, UK).
Secondary antibodies—goat anti-rabbit, goat anti-mouse (Thermo Scientific).

Biswas S., Kojonazarov B., Hadzic S., Majer M., Bajraktari G., Novoyatleva T., Ghofrani H.A., Grimminger F., Seeger W., Weissmann N., Schlossmann J, & Schermuly R.T. (2020). IRAG1 Deficient Mice Develop PKG1β Dependent Pulmonary Hypertension. Cells, 9(10), 2280.

+ Open protocol

+ Expand

Quantifying TrkB Expression in Brain Regions

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

The brains from both males and females were dissected out from adult animals (N = 4/group). The different brain structures used for analysis were the hippocampus, hypothalamus, and mid-brain. The brain samples were homogenized using the standard RIPA lysis buffer, containing a cocktail of protease inhibitors and orthovanadate, followed by centrifugation at 13,000 rpm for 15 min at + 4°C. The Rn33b cells were lysed similarly and sonicated before centrifugation. The samples were then separated using gradient gels 4–15% (NuPAGE Protein gels, Invitrogen) and blotted on to a PVDF membrane. The primary antibodies were RD-TrkB (Cat# AF397, RD systems Inc., MN, United States) and GAPDH (Cat# ab75479, Abcam). The primary antibodies were used at 1:1,000 dilution. The respective HRP conjugated secondary antibodies was blocked for 1 h at room temperature. The chemiluminescent assay was performed using ECL (Pierce). All images obtained were analyzed using ImageJ software.

Sahu M.P., Pazos-Boubeta Y., Steinzeig A., Kaurinkoski K., Palmisano M., Borowecki O., Piepponen T.P, & Castrén E. (2021). Depletion of TrkB Receptors From Adult Serotonergic Neurons Increases Brain Serotonin Levels, Enhances Energy Metabolism and Impairs Learning and Memory. Frontiers in Molecular Neuroscience, 14, 616178.

+ Open protocol

+ Expand

Western Blot Analysis of GFP

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

10 μl of cell-free samples were mixed with 2x SDS sample buffer (0.25 M Tris/HCl (pH 6.8), 4% SDS, 20% glycerol, 10% 2-mercaptoethanol, 0.25% bromophenol blue) and denaturation was performed by incubation at 95°C for 5 min. Bands were separated on NuPAGE™ protein gels (4–12%, Bis-Tris Gels by Thermo Fischer Scientific) using constant voltages (200 V for 45 min). Proteins were transferred to PVDF membranes and probed with a rabbit polyclonal anti-GFP antibody (Invitrogen, 1:2000 dilution). Anti-rabbit horseradish peroxidase-conjugated antibody was used as secondary antibody (Promega, 1:2200 dilution). The membranes were visualized by enhanced chemiluminescent kit (Thermo Fischer Scientific).

Bains J.K., Qureshi N.S., Ceylan B., Wacker A, & Schwalbe H. (2023). Cell-free transcription-translation system: a dual read-out assay to characterize riboswitch function. Nucleic Acids Research, 51(15), e82.

+ Open protocol

+ Expand

Immunoblotting Protocols for Protein Analysis

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

Immunoblotting experiments were performed by running protein lysates on NuPAGE Protein gels (Thermo Fisher) and transferring onto PVDF membranes. 4–12% Bis-Tris gels were used for data in Figs 1b and 2b, 3–8% Tris-Acetate gels were used for Figs 3c,d, and S2. Antibodies used were: MYC (Abcam #ab32072), β-actin (Sigma-Aldrich #A5441), AR (Millipore 06-680), PSA (Abcam #ab53774), BRD4 (Bethyl #A301-985A) and GAPDH (Santa Cruz #sc-32233). Blots were imaged using an Odyssey imaging system (LI-COR) according to the manufacturer’s instructions.

Coleman D.J., Gao L., Schwartzman J., Korkola J.E., Sampson D., Derrick D.S., Urrutia J., Balter A., Burchard J., King C.J., Chiotti K.E., Heiser L.M, & Alumkal J.J. (2019). Maintenance of MYC expression promotes de novo resistance to BET bromodomain inhibition in castration-resistant prostate cancer. Scientific Reports, 9, 3823.

+ Open protocol

+ Expand

Immunoprecipitation of GFP-tagged Proteins

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

Immunoprecipitation was typically carried out from close-to-confluence 10-cm dishes transfected in advance with appropriate construct. Cells were washed with PBS, collected and lysed in Net-N buffer (50 mM TrisHCl pH7.5, 1 mM EDTA, 0.5% IGEPAL CA-630, 1 mM DTT, protease inhibitors (Sigma, cOmplete mini), sonicated and cleared by centrifugation. 0.5–1 mg total protein extract was incubated with GFP_Trap beads (Chromotek) for 1 h at 4 °C with rotation. Beads were washed five times with lysate buffer and proteins were eluted by heat denaturation in loading dye. Samples were resolved on NuPAGE protein gels (ThermoFisher), transferred to nitrocellulose membrane and blocked in 5% milk. Anti-PAR (1:2000) and anti-GFP (1:2000) were incubated at 4 °C overnight. Proteins were detected and quantified on the Odyssey Fc imaging system (LiCor).

Krastev D.B., Pettitt S.J., Campbell J., Song F., Tanos B.E., Stoynov S.S., Ashworth A, & Lord C.J. (2018). Coupling bimolecular PARylation biosensors with genetic screens to identify PARylation targets. Nature Communications, 9, 2016.

+ Open protocol

+ Expand

Western Blot Analysis of p53 Protein

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

Total protein extracts (20 µg) were separated on 4% to 12% NuPAGE protein gel (ThermoFisher Scientific) and transferred to nitrocellulose membranes. The membrane was incubated with mouse monoclonal anti-human p53 DO-1 antibody (1:500; Calbiochem) and mouse monoclonal β-Actin antibody (1:4000; Sigma-Aldrich) with correspondent secondary antibodies. Membranes were imaged using a LI-COR Odyssey infrared scanner.

Quinn E.A., Maciaszek J.L., Pinto E.M., Phillips A.H., Berdy D., Khandwala M., Upadhyaya S.A., Zambetti G.P., Kriwacki R.W., Ellison D.W., Nichols K.E, & Kesserwan C. (2019). From uncertainty to pathogenicity: clinical and functional interrogation of a rare TP53 in-frame deletion. Cold Spring Harbor Molecular Case Studies, 5(4), a003921.

+ Open protocol

+ Expand

Affinity Purification of RAD51AP1 Complexes

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

MBP- and His₆-tagged full-length RAD51AP1 or MBP- and His₆-tagged RAD51AP1-F1, -F2, -F3 (80 nM each) were incubated with pre-equilibrated Ni-NTA resin (Thermo Fisher Scientific) in buffer F (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Triton X-100, 2% BSA) at 4 °C with gentle rotation for 1 h. The supernatant was removed and bound protein was further incubated with histone Octamer (80 nM) in buffer F containing DNase (1 U/μg protein) and at 4 °C with gentle rotation for 2 h. The resin then was washed 4 × with buffer F and bound protein complexes were eluted in 40 μl buffer F with 300 mM imidazole. Eluted complexes (10 μl) were fractionated on a 10% NuPAGE protein gel (Thermofisher Scientific), transferred onto a PVDF membrane, and detected by western blot analysis.

Pires E., Sharma N., Selemenakis P., Wu B., Huang Y., Alimbetov D.S., Zhao W, & Wiese C. (2021). RAD51AP1 mediates RAD51 activity through nucleosome interaction. The Journal of Biological Chemistry, 297(1), 100844.

+ Open protocol

+ Expand

SDS-PAGE Analysis of Recombinant Proteins

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

Recombinant proteins were analyzed by SDS–PAGE analysis to determine recombinant protein size and integrity. The recombinant protein (15 μg) was mixed with Tris–glycine SDS sample buffer (Thermo Fisher, Rockford, IL, USA), heated at 70°C for 10 min, and then loaded onto a NuPAGE protein gel (Thermo Fisher, Rockford, IL, USA). Staining and de-staining of the gel were performed with eStain L1C protein staining system (GenScript, Piscataway, NJ, USA).

Bold D., Roman-Sosa G., Gaudreault N.N., Zayat B., Pogranichniy R.M, & Richt J.A. (2022). Development of an Indirect ELISA for the Detection of SARS-CoV-2 Antibodies in Cats. Frontiers in Veterinary Science, 9, 864884.

+ Open protocol

+ Expand

RAD51AP1 Protein-Histone Binding Assay

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

MBP-and His6-tagged full-length RAD51AP1 or MBP-and His6-tagged RAD51AP1-F1, -F2, -F3 (80 nM each) were incubated with pre-equilibrated Ni-NTA resin (Thermofisher Scientific) in buffer F (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Triton X-100, 2% BSA) at 4°C with gentle rotation for 1 h. The supernatant was removed and bound protein was further incubated with histone Octamer (80 nM) in buffer F containing DNase (1 U/µg protein) and at 4°C with gentle rotation for 2 h. The resin then was washed 4´ with buffer F and bound protein complexes were eluted in 40 µl buffer F with 300 mM imidazole. Eluted complexes (10 µl) were fractionated on a 10% NuPAGE protein gel (Thermofisher Scientific), transferred onto a PVDF membrane and detected by Western blot analysis.

Pires E., Sharma N., Selemenakis P., Wu B., Huang Y., Alimbetov D., Zhao W., & Wiese C. (2020). RAD51AP1 mediates RAD51 activity through nucleosome interaction.

+ 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!