b'NuPAGE Tris-Acetate gelsHigh molecular weight protein separation Tris-acetate gel chemistry enables the optimal separationSpecificationsof high molecular weight proteins. NuPAGE Tris-Acetate Shelf life: 8 monthsgels offer a pH 8.1 environment that minimizes protein modifications and results in sharper bands. NuPAGE Average run time: 35 minutesTris-Acetate gels can also be run with Novex Tris-Glycine Separation range: 30400 kDaNative Running Buffer to resolve native proteins more effectively than a Tris-glycine gel system. Polyacrylamide concentrations: fixed 7%; gradient 38%NuPAGE Tris-Acetate gels and buffers are designed Gel dimensions:to allow: Mini: 8 x 8 cm (1 or 1.5 mm thick) Optimal separation of high molecular weight proteins Midi: 8 x 13 cm (1 mm thick)(Figures 13 and 15) Maximum sample volume per 10-well mini gel: Preservation of protein sample integrity using optimized25 L (1 mm thick); 37 L (1.5 mm thick)sample preparation processesMore efficient western blot transferneutral pH prevents reoxidation of reduced samples during protein transfer (Figure 14)20 g 6.7 g 2.3 g 0.74 g 0.25 g 83 ng 28 ng 9 ngesults acquired witR hEGFR (~190 kDa)the Miniank NuPAGE 38% Tris-Acetate mini gelGel T20 g 10 g 5 g 2.5 g 1.25 g 0.75 g 0.62 gEGFR (~190 kDa)Novex 420% Tris-Glycine mini gel, WedgeWell formatFigure 13. NuPAGE Tris-Acetate gel electrophoresis. Protein standardsFigure 14. Improved transfers of high molecular weight proteins and samples were loaded at 10 L sample volumes in an Invitrogenenhance western detection sensitivity. Western blotting analysis of NuPAGE 38% Tris-Acetate gel. Electrophoresis was performedEGFR from A431 lysates transferred from a Novex 420% Tris-Glycine mini using the Mini Gel Tank at 200 V (constant). Sharp, straight bandsgel, WedgeWell format, and a NuPAGE 38% Tris-Acetate mini gel using were observed after staining with SimplyBlue SafeStain. Images werethe iBlot 2 Gel Transfer Device.acquired using a flatbed scanner. Lane 1: SeeBlue Plus2 Prestained Standard; lane 2: 10 g E. coli lysate; lane 3: Mark12 Unstained Standard (blend of 12 purified proteins); lane 4: 40 g HeLa cell lysate; lane 5: 20 g HeLa cell lysate; lane 6: 5 g BSA; lane 7: 40 g Jurkat cell lysate; lane 8: 5 g GST fusion protein; lane 9: Novex Sharp Unstained Protein Standard; lane 10: 5 g -galactosidase.22'