蛋白质技术

Transfer BufferBjerrum and Schafer-Nielsen transfer buffer containing SDS:48 mM Trizma base 39 mM glycine 20% methanol 0.00375% SDSDissolve 5.82 g of Trizma base and 2.93 g of glycine in about 700 ml ...

This is a protocol using the glycine system described by Laemmli U.K. (1970) Nature 227:680-685. For polypeptides smaller than 30 kDa the tricine system described by Schagger H. & Von Jagon G. (1987) ...

MaterialsPolyacrylamide pre-cast gel SimplyBlue™ Coomassie protein stain 1x pre-mixed solution (Coomassie blue dye binds to proteins (arginine the aromatic amino acids and histidine) allowing protein ...

Materials Densitometer 17 Washed and stained gel from Exercise 4.2 Procedure Using the scanning cuvette supplied with your densitometer/scanner calibrate the scanner and choose the proper scanning mod ...

Make up following solutions (maybe stored for months at room temp.) Ratios are v/v unless stated otherwise. Stain:40% methanol; 10% acetic acid; 50% water; 0.1 % (w/v) Coomassie Brilliant Blue R250De- ...

LEVEL I Materials Protein gel from Exercise 4.2 0.25% (w/v) Coomasie Brilliant Blue R 250 in methanol-water-glacial acetic acid (5-5-1) filtered immediately before use. 7% (v/v) acetic acid Commercial ...

Silver Stain is about 10 to 100 times more sensitive than Coomasie Blue Staining; at least 1ng protein using this protocol. Estimation of protein amouts is not advisable since the staining intensity d ...

IntroductionSilver staining is the most sensitive protein staining method available for gel electrophoresis so far. It can detect as little as 5 ng protein in a 2.5 mm wide band in 0.75 mm thick gel. ...

We currently use the SilverQuest Silver stain kit from Invitrogen. This method is faster than the method listed below. The rapid method works well for most applications. Click here for instructions to ...

Make up following solutions: (maybe stored for months at room temp.) Ratios are v/v.Fixative: 40% methanol; 10% acetic acid; 50% waterWash Sol.:30% ethanol in waterWater:d.i. waterReductant:200 mg sod ...

Inc. Time1. Gel Fix Solution (50% MeOH 12% TCA 2% CuCl2)20 min. 100% MeOH250ml (Good for 1 month) 100% TCA60ml CuCl210g H2Oto 500ml 2. Solution A (10% EtOH 5% Acetic Acid)10 min. 100% EtOH100ml ...

Immediately after the electrophoretic run is terminated slice the stacking gel from the slab and place the gel in a glass dish with 200 ml (8mm x 7mm mini-gel) of 40% methanol/10% acetic acid for the ...

Materials Protein gel from Exercise 4.2 45% (v/v) Methanol + 12% (w/v) acetic acid 5% (v/v) Methanol + 7% (w/v) acetic acid 10% Glutaraldehyde 0.01M Dithiothreitol Silver nitrate solution Sodium citra ...

This material is accompanied by a presentation on protein structure and principles behind denaturing samples and discontinuous gel electrophoresis.The separation of macromolecules in an electric field ...

Assemble each gel sandwich by stacking in order the notched aluminum plate two 0.75-mm spacers and a glass plate. It is important that the spacers are aligned properly with the ends flush with the top ...

Stock Solutions1) 1.5M TrisHCl pH 8.3 + 0.4% SDS (adjust pH before you add the SDS). Keep RT. 2) 30% Acrylamide 0.8% Methylene bis Acrylamide or 40% Acrylamide / Methylene bis Acrylamide (ratio: 37.5: ...

You will need the following reagents:5x Sample Buffer10% w/vSDS10 mM Dithiothreitol or beta-mercapto-ethanol20 % v/v Glycerol0.2 M Tris-HCl pH 6.80.05% w/vBromophenolblueShould add up to 8M urea for r ...

Stock Solutions1) 1.5M TrisHCl pH 8.9 - Keep RT. 2) 30% Acrylamide 0.8% Methylene bis Acrylamide. Keep 4°C. 3) 0.5M TrisHCl pH 6.8 - Keep RT. 4) 10% Ammonium Persulfate (APS). Keep 4°C less than 1 mon ...

USE ONLY FRESH GELSStock Solutions1) 1.5M Acetate-KOH pH 4.3 (48ml 1MKOH + 17.2ml AcH + H2O up to 200ml) - Keep RT. 2) 30% Acrylamide 0.8% Methylene bis Acrylamide. Keep 4°C. 3) 0.25M Acetate-KOH pH 6 ...

Assemble each gel sandwich by stacking in order the notched aluminum plate two 0.75-mm spacers and a glass plate. It is important that the spacers are aligned properly. Fit the gel sandwiches (usually ...