Big Chemical Encyclopedia

Chemical substances, components, reactions, process design ...

Articles Figures Tables About

Coomassie blue solution preparation

After completion of electrophoresis, the gel was stained with Coomassie blue. To prepare the staining solution, 40 mg Coomassie blue R-250 was dissolved in 25 ml isopropanol and 10 ml glacial acetic acid. The solution was filtered and the volume was increased to 100 ml with distilled water. The gel was placed in staining solution for 1 h, followed by washing with 10% acetic acid/2.5% isopropanol. [Pg.111]

Prepare a solution of Coomassie Blue as follows (Bradford, 1976) ... [Pg.334]

The Coomassie staining solution is prepared as follows add 40 mL of 85% o-phosphoric acid, 40 g of (NH4)2S04, and 0.48 g of Coomassie Blue G-250 to 360 mL deionized water. Stir solution and add 100 mL methanol. [Pg.29]

Separate the sandwich after 8 min. Place the gel into 25 ml of gel fixing solution and prepare for staining with Coomassie blue or with silver. Place the film into a dry petri dish (for best contrast on a light box) and slowly pour 30 ml of developing buffer into the dish. Do not agitate the dish. Temperature and pH of the buffer depend on type and amount of the protease investigated and should reflect optimal reaction conditions. [Pg.267]

Separation and Purification of IgG via SDS-PAGE and Electrotransfer of Immunoglobulines onto an Inert PVDF Membrane About 50 pg of a glycoprotein is mixed with the same volume of a double concentrated SDS buffer and denatured at 100 °C. The samples are applied onto the prepared SDS polyacrylamide gel, concentrated electrophoretically in the collecting gel, and separated to piuity in the separation gel. After this step, the separation gel is put onto a previously activated PVDF membrane (Immobilon PSQ, 0.1 pm pore size, Millipore, USA) and enclosed in a Western Blot cassette. After electrotransfer, the protein on the membrane is stained with a 0.1% Coomassie Brilliant Blue solution. The membrane is washed several times to remove residues from the SDS gel electrophoresis. The completeness of the transfer can be checked by staining the gel in the... [Pg.1362]

Protein concentration can be determined using a method introduced by Bradford,4 which utilises Pierce reagent 23200 (Piece Chemical Company, Rockford, IL, USA) in combination with an acidic Coomassie Brilliant Blue G-250 solution to absorb at 595 nm when the reagent binds to the protein. A 20 mg/1 bovine serum albumin (Piece Chemical Company, Rockford, IL, USA) solution will be used to prepare a standard calibration curve for determination of protein concentration. The sample for analysis of SCP is initially homogenised or vibrated in a sonic system to break down the cell walls. [Pg.16]

Prepare staining solution by dissolving 1.25 g coomassie brilliant blue in a mixture of 227 ml distilled water, 227 ml methanol, and 46 ml glacial acetic acid (5 5 1). When the dye is fully dissolved, filter the solution through Whatman No. 1 filter paper or its equivalent. [Pg.220]

The reagent is prepared as follows. Coomassie brilliant blue C-250 (100 mg) is dissolved in 50 cm 95% ethanol. To this solution phosphoric acid (100 cm, 85% w/v) is added and the solution diluted to 1 dm. To perform the assay, x cm of the sample containing 5-100 J,g of protein is placed in a clean, dry test tube. (0.5 - x) cm water and 5.0 cm of diluted dye reagent are added and the solution mixed well. After a period of from 5-60 min, A595 is determined. [Pg.138]

Protein determination measurements were performed by Bradford s Method. Bradford reagent was prepared by mixing 25 ml phosphoric acid, 12.5 ml ethanol and 25 mg Coomassie Brilliant Blue (G-dye). The mixture was diluted to 50 ml with distilled water. During measurements, a solution of Bradford reagent was prepared by mixing 1 volume stock solution with 4 volumes of distilled water. [Pg.160]


See other pages where Coomassie blue solution preparation is mentioned: [Pg.916]    [Pg.530]    [Pg.188]    [Pg.537]    [Pg.553]    [Pg.206]    [Pg.119]    [Pg.213]    [Pg.537]    [Pg.553]    [Pg.265]    [Pg.157]    [Pg.3260]    [Pg.388]    [Pg.317]    [Pg.317]    [Pg.357]    [Pg.76]    [Pg.171]    [Pg.183]    [Pg.159]    [Pg.263]    [Pg.79]   


SEARCH



Blue solutions

Coomassie

Coomassie blue

Solution preparing

© 2024 chempedia.info