The Mini-PROTEAN Tetra Cell can be configured for use with 1–4 precast or handcast gels in different thicknesses. Requires a PowerPac Basic or comparable power supply.Leak-free electrophoresis and gel casting.Indestructible molded polycarbonate construction.Runs 1–4 mini polyacrylamide gels in 30 min.
BIO RAD WESTERN BLOT GEL SERIES
Protein Expression and Purification Series.Comparative Proteomics Kit I: Protein Profiler Module.
Use the Mini-PROTEAN Tetra cell to teach protein fingerprinting, sample purity analysis, and downstream western blotting techniques in independent projects or with the following educational kits: An indispensable piece of equipment in research laboratories, the durable and easy-to-assemble Mini PROTEAN Tetra cell is also well-suited for use in classroom laboratories. Prepare 0.Bio-Rad’s Mini-PROTEAN Tetra cell is a mini-format, vertical polyacrylamide gel electrophoresis system that provides rapid, high-resolution separation of protein (or DNA) mixtures by SDS-PAGE or native PAGE. Mix the Clarity Western ECL Substrate Kit components in a 1:1 ratio.
It is important to use an ECL substrate that has good sensitivity and long signal duration, such as the Clarity Western ECL Substrate. After the final wash step, keep the blot in TBST while preparing for blot detectionĪll PrecisionAb Antibodies were validated using enhanced chemiluminescent (ECL) detection. Rinse the blot with 15 ml TBST at RT for 5 min. 6.2 DNA Blotting (For acrylamide gels with DNA 250 bp to 1 kb). Incubate the blot in the secondary antibody and blocking buffer solution at RT for 1 hr with gentle agitation Bio-Rad also recommends that all Trans-Blot SD cell components and accessories be. Please refer to the antibody product page for details on the exact secondary antibody used during the validation process. Repeat for a total of five washesĭilute the appropriate secondary antibody in 10 ml blocking buffer according to the following table: Scrape adherent cells off the dish using a cold plastic cell scraper and gently transfer the cell suspension into a precooled microcentrifuge tube.
Aspirate the TBS, then add ice-cold RIPA buffer (1 ml per 100 mm dish). ERAP2 Allelic Variants Analyses 199 Cells 2020, 9, 1951 3.2. Incubate the blot in the primary antibody and blocking buffer solution at 4☌ overnight with gentle agitation Place the cell culture dish in ice and wash the cells with ice-cold Tris-buffered saline (TBS). Please see the validation protocol (bulletin 6603) for more details.ĭilute the primary antibody 1:1,000 in 10 ml blocking buffer There are a wide selection of Mini-PROTEAN Combs, available in 0.75 mm, 1.0 mm, and 1. If using BSA, you may notice some nonspecific bands due to its low stringency. Choose these Mini-PROTEAN 15-well combs for use with spacer plates with integrated 1.5 mm spacers for preparing gels with the Mini-PROTEAN Tetra Handcast Systems and Mini-PROTEAN 3 Multi-Casting Chamber.Packaging Options. We recommend using casein or nonfat dried milk for blocking. When using casein, do not block for longer than 30 min to prevent reduction in signal specificity. Load the control cell lysate adjacent to your samples and the molecular weight (MW) marker (see diagram).ĭuring the validation process, we blocked for 30 min at room temperature (RT) in blocking buffer + 0.1% Tween 20. If using BME, add 180 μl H20, 200 μl 2x Laemmli Sample Buffer, and 20 μl BME If using DTT, add 190 μl H20, 200 μl 2x Laemmli Sample Buffer, and 10 μl 2 M DTT Reconstitute 400 µg lysate in one of the following ways, depending on the reducing reagent used:
Secondary antibodies (see antibody datasheet)
BIO RAD WESTERN BLOT GEL PLUS
Precision Plus Protein All Blue Standards Value Pack.Mini-PROTEAN Tetra Cell for Mini Precast Gels.Any kD Mini-PROTEAN® TGX Stain-Free Precast Gels (10 well, 50 µl) Introduce students to protein extraction, quantitation, separation, and analysis using techniques such as polyacrylamide gel electrophoresis (PAGE), western.4–15% Mini-PROTEAN® TGX Stain-Free Precast Gels (10 well, 50 µl).Phosphate Buffered Saline (PBS) containing 1% w/v BSA Reducing agents such as dithiothreitol (DTT) or ß-mercaptoethanol (BME)Įlectrophoresis gels, reagents, and equipment Quantitate amount of protein using Bio-rad Dc Protein Asssay (similar to Bradford). Use the charts below to determine the best gel type and percentage for SDS-PAGE and other protein electrophoresis applications.
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