Related Manuals for Nippon Genetics Europe FastGene PG01
Summary of Contents for Nippon Genetics Europe FastGene PG01
- Page 1 PAGE PROTEIN SYSTEM PG01 Manual NIPPON Genetics EUROPE GmbH Mariaweilerstraße 28-30, 52349 Düren, Germany, www.nippongenetics.eu Tel: +49 2421 5549 60, Fax: +49 2421 55496 11, E-Mail: info@nippongenetics.de,...
- Page 2 FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 3: Table Of Contents
3.2.3 PAGE System Chamber Assembly........P. 23 3.2.4 Sample Preparation.............. P. 24 3.2.5 Sample Loading..............3.2.6 Performing Electrophoresis..........P. 25 4. TROUBLE SHOOTING..................P. 27 5. WARRANTY......................P. 31 P. 31 6. CONTACT INFORMATION................FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 4 FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 5: Introduction
After the PAGE protein separation is completed, protein visualization and detection is carried out. Protein visualization is achieved by the use of protein specific stains. After staining, images can be taken of the gel for analytical purposes. FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 6 FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 7: System Components
Large gel electrophoresis buffer tank ® (1 chamber tank) PG06 FastGene Gel casting clip Frame for holding hand-cast gels (4 pieces) ® PG07 FastGene Sealing gaskets Sealing gaskets for gel casting (5 pieces) ® FastGene PAGE Protein System ® NIPPON Genetics EUROPE... -
Page 8: Page System Set Components
FastGene PAGE Protein System. If you find there is something wrong with the ® system or the packaging content, please contact NIPPON Genetics EUROPE or your local distributor. The FastGene PAGE Protein System (PG01) contains PAGE System components and Gel ®... - Page 9 4. Inner electrophoresis chamber 5. Hand-cast gel / 6. U sealing strip without electrodes (with side clip Pre-cast gel / for gel holder 2x) Plastic dummy cassette Fig. 1: PAGE system components FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
- Page 10 (cathode) from the (outer) gel/blot chamber tank (anode). Depending on the gel size and gel type, different U sealing strips are used (See page 19 for the use of compatible U sealing strips). FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 11: Gel Hand-Casting Set Components
Sealing gaskets (PG07) Glass plate holder Tube holder Glass plate short Glass spacer long 1 mm (0.75 mm, 1.5 mm) Gel casting base Gel casting clip Sealing gasket Fig. 2: Gel hand-casting set components FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 12 FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 13: System Operation
Place the gel casting clip on a horizontal lab desk and keep both hinges of the frame open. Position the short glass plate towards the front and slide the assembled glass spacer and glass plate into the gel casting clip from the top. FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 14 Carefully check that both glass plates are level with each other and are aligned at the bottom of the gel casting clip. Adjust if necessary. A wrong alignment can lead to leakage during gel casting. FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
- Page 15 (1 mm, 0.75 mm, 1.5 mm) Spring leverage Continue with polyacrylamide gel casting. Repeat steps 1-6 when casting more than one gel. FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 16: Polyacrylamide Gel Casting
0,1 % 10 % APS 30 µl 75 µl 75 µl 0,05 % Total Volume 6 ml 15 ml 15 ml *Add later, right before gel casting (step 5 and step 7) FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 17 Remove the comb again. Glass plate short Comb (e.g. 1 mm, 15 well) (in front) Glass spacer long 1 mm (0.75 mm, 1.5 mm) Mark: ~ 1 cm below teeth of comb FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
- Page 18 For optimal results, only use high quality reagents. Always prepare fresh APS solutions and avoid using TEMED solutions that are older than three months. • Do not store gels for a long period of time. Do not freeze the gels. FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 19: Page System Operation
• 10 x 10 cm gels • FastGene Pre-cast gels • Bio-Rad TGX • ThermoFisher mini ® • Hand-cast gels Fig. 4: U sealing strips compatibility with Pre-cast gels and Hand-cast gels FastGene PAGE Protein System ® NIPPON Genetics EUROPE... -
Page 20: Gel Cassette Assembly
Inner electrophoresis chamber. Rest the lower part of a gel cassette on the two steps positioned at the bottom part of the inner electrophoresis chamber and move the upper part of the gel cassette towards the U sealing strip. FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 21 The gel cassette now rests on the two moveable knobs and is pressed upwards. This additionally secures the glass plates and prevents buffer leakage. Side clip fastener Side clip fastener (open) (closed) FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 22: Page System Chamber Assembly
Fill the outer Gel/blot chamber tank up to the indicator mark for 2 Gels (using one inner electrophoresis chamber) or 4 Gels (using two inner electrophoresis chambers). FastGene PAGE Protein System ® NIPPON Genetics EUROPE... -
Page 23: Sample Preparation
The disruption of disulfide bonds is carried out by the addition of reducing agents such as DTT (dithiothreitol) or b-ME (beta-mercaptoethanol). FastGene PAGE Protein System ® NIPPON Genetics EUROPE... -
Page 24: Sample Loading
Wash out the gel wells (also wells that will be kept empty) with running buffer before loading the samples to ensure an even buffer concentration in the wells and avoiding "smile" effects. FastGene PAGE Protein System ® NIPPON Genetics EUROPE... -
Page 25: Performing Electrophoresis
Do not reuse the electrophoresis running buffer. Discard the running buffer and clean the inner electrophoresis chambers, U sealing strips and the Gel/blot chamber tank with distilled water. FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 26 FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 27: Trouble Shooting
(APS and TEMED) by 20 % Inhomogenous gel, Catalyst not active Use a fresh catalyst solution • • • polymerization too (APS, TEMED) slow Increase catalyst • concentration by 20 % FastGene PAGE Protein System ® NIPPON Genetics EUROPE... - Page 28 Wrong running buffer Check buffer concentration and • • • electrophoresis concentration make a new running buffer runs too fast Voltage too high Decrease voltage by 50 % • • FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
- Page 29 Fill the Inner electrophoresis • • chamber completely with running buffer, make sure the wells are covered with running buffer Different buffer Wash out gel wells with running • • concentration in wells buffer FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
- Page 30 No protein Wrong charge of Check pH of stacking gel, • • • migration in migrating protein resolving gel and running gel resolving gel (neutral or positive) buffer FastGene PAGE Protein System ® NIPPON Genetics EUROPE...
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Page 31: Warranty
Genetics EUROPE's liability under the warranty is limited to the receipt of adequate evidence by the customer that the defect falls under the warranty conditions. All claims under this warranty must be submitted to NIPPON Genetics EUROPE within one year of delivery of the product to the customer. - Page 32 NIPPON Genetics EUROPE GmbH Mariaweilerstraße 28-30, 52349 Düren, Germany, www.nippongenetics.eu Tel: +49 2421 554960 Fax: +49 2421 55496 11, E-Mail: info@nippongenetics.de,...