Nupage novex bis tris protein gels

Manufactured by Thermo Fisher Scientific

The NuPAGE® Novex® Bis-Tris Protein Gels are pre-cast polyacrylamide gels designed for the separation of proteins using a Bis-Tris buffering system. They are suitable for a wide range of molecular weights and can be used in various electrophoresis techniques.

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Lab products found in correlation

Page blue protein stain, by Thermo Fisher Scientific (1 mentions) High performance chemiluminescence film, by GE Healthcare (1 mentions) Trans blot sd semi dry transfer cell, by Bio-Rad (1 mentions) Micro bio spin column, by Bio-Rad (1 mentions) Bio safe coomassie, by Bio-Rad (1 mentions) Iblot, by Thermo Fisher Scientific (1 mentions) Sigmafast bcip nbt, by Merck Group (1 mentions)

Close spelling variants of this product

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3 protocols using nupage novex bis tris protein gels

Immunoblot Analysis of Adhesins

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Bacterial samples from at least five independent experiments, were collected from plate, washed twice in Sia buffer. SDS Page analysis was made using NuPage Novex Bis-Tris protein gels (4–12% or 12%) using 1x MOPS SDS as running buffer (Life Technologies). The gels were either stained with PageBlue Protein stain (Thermo Scientific) or transferred to PVDF membrane using Trans-Blot SD Semi-Dry Transfer Cell (Bio Rad). Immunoblot analysis was performed as previously described44 (link). Antibodies against AlpB (AK262)45 (link), SabA (AK278) and BabA (AK277), were used in combination with secondary anti-rabbit IgG-HRP (P0160, DAKO A/S, Denmark). Blots were developed with SuperSignal West Pico Chemiluminescent Substrate (Pierce, Rockford, IL) and detected on High Performance Chemiluminescence film (GE Healthcare). BabA, SabA and AlpB protein densities were measured by ImageJ software (a public domain image processing program from the National Institutes of Health, Maryland, USA).The adhesin/AlpB density ratio for each sample was used to calculate the fold difference between the wt and the arsS deletion mutant.

Skoog E.C., Padra M., Åberg A., Gideonsson P., Obi I., Quintana-Hayashi M.P., Arnqvist A, & Lindén S.K. (2017). BabA dependent binding of Helicobacter pylori to human gastric mucins cause aggregation that inhibits proliferation and is regulated via ArsS. Scientific Reports, 7, 40656.

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Immunoprecipitation of Familial Hypercholesterolemia Protein

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To isolate FH from a small amount of sample an IP micro-method was developed using 5H5-Sepharose resin. The final standard IP protocol consisted in incubation of 50 µl of serum with 100 µl of 5H5-sepharose resin (1:2 slurry in PBS) in a total volume of 500 µl of PBS, followed by washes in PBS/TW and elution in 0.1 M Glycine pH 3.0. The tubes were centrifuged at each step at 10,000 x g for 30 sec. to precipitate the resin and aspirate the supernatant. The method was further improved by the use of Micro Bio-Spin™ Columns (Bio-Rad) which allow for a faster procedure.
The protocol was essentially the same except that the samples were collected from the bottom of the micro-columns. Samples were analysed by SDS-PAGE on 4-12% NuPAGE® Novex® Bis-Tris Protein Gels (Life Technologies) in MOPS 1X buffer at 200 V for 60 min, followed by Coomassie staining with BioSafe Coomassie (Bio-Rad).

Berra S, & Clivio A. (2016). Rapid isolation of pure Complement Factor H from serum for functional studies by the use of a monoclonal antibody that discriminates FH from all the other isoforms. Molecular immunology, 72.

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Western Blot Protein Visualization

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Samples were run on 4-12% NuPAGE® Novex® Bis-Tris Protein Gels (Life Technologies) in MOPS 1X buffer at 200 V for 60 min.
Gels were blotted on a PVDF membrane using iBLOT (Life Technologies) with protocol P3 for 7-10 min. Membranes were blocked overnight in TBST with 3% BSA at 4°C. Primary antibodies were typically diluted in TBST with 1% BSA and applied to the membranes 1h RT in constant agitation. Membranes were then washed 3 times with TBST and incubated 1h RT in constant agitation with the corresponding alkaline phosphatase conjugated secondary antibody diluted in TBST with 1% BSA. After 3 washes in TBST, specific signal was revealed with SigmaFast™ BCIP/NBT (Sigma-Aldrich).

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