SE245 User Manual
Contents
1. The screws face outward Seal the sandwich by turning both cams so that the short end of the cam the arrow points down Dual Gel Caster Important The plate and spacer bottoms must be aligned for a proper seal 6 p4 Caution Acrylamide is a neurotoxin Always wear gloves and observe all laboratory safety procedures Pour the gel 0 Prepare the required amount of monomer solution Deaerate and add the initiator and catalyst just prior to pouring the gel Use a pipet to deliver the solution into one corner of the plate taking care not to intro duce any air pockets See below for the appropriate solution level according to the application No stacking gel Fill solution to just below the top of the notched plate If air pockets form remove with a pipet or syringe Introduce a comb at a slight angle into each sandwich taking care not to trap air bubbles under the teeth Stacking gel Fill solution to 3 cm below the top of the rectangular glass plate This height allows 1 cm of stacking gel below the wells Pour the gel and apply an overlay After the gel is polymerized prepare the stacking gel as described below 2 D electrophoresis Fill solution to about 1 5 cm below the top of the rectangular glass plate and apply an overlay This height allows 4 5 mm of space for the IPG strip and an agarose seal 9 Overlay each gel with a thin layer of water saturated n butanol water or2. 75 1 00 10 5 2 SE2619T 2 1 0 1 50 10 5 2 SE2619T 2 1 5 Combs no of wells thickness mm width mm 5 0 75 13 0 1 SE211A 5 75 5 1 00 13 0 1 SE211A 5 1 0 5 1 50 13 0 1 SE211A 5 1 5 9 1 00 5 8 1 SE211A 9 1 0 10 0 75 4 8 1 SE211A 10 75 10 1 00 4 8 1 SE211A 10 1 0 10 1 50 4 8 1 SE211A 10 1 5 12 1 00 4 75 1 SE211A 12 1 0 15 0 75 2 9 1 SE211A 15 75 15 1 00 2 9 1 SE211A 15 1 0 15 1 50 2 9 1 SE211A 15 1 5 18 1 00 2 9 1 SE211A 18 1 0 1 1 0 75 68 5 1 SE211A R 75 1 1 1 00 68 5 1 SE211A R 1 0 1 1 1 50 68 5 1 SE211A R 1 5 Microtiter spacing Preparative reference well e p10 Hoefer Inc 84 October Hill Road Holliston Toll Free A 01746 800 227 4750 Phone 1 508 893 8999 Fax 1 508 893 0176 E mail su Web www pport hoeferinc com hoeferinc com Hoefer is a registered trademark of Hoefer RBS 35 is nc a trademark of Pierce Chemical Co 2012 Hoefer Inc All rights reserved Printed in the USA Hoefer
3. diluted gel buffer to minimize exposure of the top surface of the gel solution to oxygen Deliver the overlay solution slowly from a glass syringe fitted with a 22 gauge needle Apply the solution near the spacer and allow it to flow across the surface unaided Allow the gel to polymerize for a minimum of one hour Stacking gel preparation Pour a stacking gel either while the sandwich is still in the gel caster or after it is transferred to the electrophoresis unit see instrument instruc tions Resolution is optimal when the stacking gel is poured just before electrophoresis To pour a stacking gel in the dual gel caster 0 Remove the overlay by rinsing the top of the gel several times with distilled water Invert the caster to drain To ensure a seamless contact between the separating and stacking gels remove residual liquid by blotting one corner with a lint free tissue 2 Calculate the volume of stacking gel monomer solu tion required Prepare the stacking gel monomer solution deaerate it and add catalyst and initiator Pour the stacking gel onto the resolving gel with a Pasteur pipet Q Introduce a comb at a slight angle into the sand wich taking care not to trap air under the teeth Allow the gel to polymerize After polymerization Oo To remove combs if necessary cover the top of the gel vvith 1X separating or stacking gel buffer then work each comb out slowly
4. Q rana SE245 Dual Gel Caster um SE245 IM Rev 10 08 12 A Ato e fe r Page finder Dual Gel Caster function and description 1 UNpackins aio e 1 Prepare the gel caster ana nee eee neneve nenes 2 Construct the gel sandwich stack and place it into the caster aaa anime a coz 2 POUR EAS pel sat ves s ide sosht h r b Stacking gel preparation kana nana nana eneve 6 After polymerizatl on adsa nai ie 7 Troubleshooting cs icactenediscndasadegatsvantiseanssaacdanscadabanacs 8 Care and Maintenance 8 Ordering information i 9 epi Fig 1 Dual Gel Caster The gel sandwich is held together by the casting clamp assembly Dual Gel Caster function and description The SE245 Dual Gel Caster holds one or two glass or glass alumina plate gel sandwiches for casting acrylamide gels A cam seals the bottom of the sandwich against a rubber gasket Once the gel is formed sandwiches are transferred to a Hoefer mini vertical unit for electrophoresis Unpacking Unwrap all packages carefully and compare contents with the packing list making sure all items arrived If any part is missing contact your local sales office Inspect all components for damage that may have occurred while the unit was in transit If any part appears damaged contact the carrier immediately Be sure to keep all packing material for damage claims or to use should it become necessary
5. by gently rocking it side to side while pulling it out 124 Remove the gel sandwich from the casting cradle by loosening the pressure bar screws tilting the sand wich forward and lifting it out Rinse the sandwich with distilled water to remove the buffer and extra gel then blot the sandwich with a lint free tissue Q To run gels Follow the instructions accompanying the electrophoresis unit To store gels Wrap individual gels in plastic wrap after adding approx 5 ml of 1X separating gel buffer to the top of each sandwich Alternatively lay a batch of gels in 1X separating buffer Store gels at 4 C for up to one week Important Decontaminate instrument of all radioactivity and or infectious agents before returning item Include documentation to verify that this has been done Troubleshooting Gel sandwich leaks e Replace any chipped plates e Check plate and spacer alignment and realign if necessary e Check the black gasket for cuts or cracks and replace if necessary e Apply a light film of Gel Seal to the bottom outside corner of each spacer Sample wells damaged or irregular e Remove air pockets and bubbles before inserting combs Slide comb into solution at an angle If bubbles persist remove comb add more monomer solution and reinsert the comb e Allow a minimum of 1 hour for polymerization of acrylamide gels e Remove the comb at a slight angle and very slowly to prevent
6. damage to the gel Care and maintenance e Thoroughly wash and rinse all caster components immediately after use e Do not autoclave or heat any part above 45 C e Do not use organic solvents abrasives strong cleaning solutions or strong acids or bases to clean any plastic part e Do not soak the gaskets Clean with a mild deter gent and allow to air dry Clean glass plates and spacers with a dilute solution of a laboratory detergent such as RBS 35 Pierce Chemical Co then rinse thoroughly with tap and distilled water Glass plates can also be treated with but not stored in acid cleaning solutions Ordering information product quantity code no Gel Caster and replacement parts SE245 Dual Gel Caster complete 1 SE245 For 1 or 2 gels 10 x 8 10 5 Sealing gasket set 2 SE246 Casting clamp assembly 1 SE249 Cams black 4 SE6005L Clamps red 4 SE252 Gel Seal 1 4 oz 1 SE6070 Glass and alumina plates 10 x 8 cm for SE250 Notched alumina plate 1 SE202N Notched alumina plates 10 SE202N 10 Rectangular glass plates 10 SE202P 10 10 x 10 5 cm for SE260 and miniVE Notched alumina plates 5 SE262N 5 Notched glass plates 5 SE262GN 5 Rectangular glass plates 5 SE262P 5 Spacers thickness length mm cm quantity code no For SE250 0 75 8 2 SE2119T 2 75 1 00 8 2 SE2119T 2 1 0 1 50 8 2 SE2119T 2 1 5 For SE260 and miniVE 0 75 10 5 2 SE2619T 2
7. lates Fig 2 Alternatively assemble the sandwich while it is in a standing position align the plate bottoms on a flat surface and align the spacers with the sides of the plates Fig 3 ESSO Note Wear gloves to keep the caster and plates free of finger marks Slide the sandwich into the casting clamp assem bly notched plate facing the back block and plate bottoms flush with the flat surface Q While holding the sandwich in place secure it by tightening all 6 screws until they are finger tight To prevent breaking plates do not over tighten Inspect plate and spacer alignment To prevent leak ing the bottom of the plates and spacers must be aligned and the sandwich should protrude slightly below the back block Adjust if necessary 6 Place the white foam gasket into the bottom of the casting slot and lay the black gasket over it Oo Place the clamp assembly in the casting cradle screw side facing out In this position the gel will be visible through the rectangular glass plate 18 Insert a cam into each hole on both sides of the casting cradle with the arrow short end pointing up Seal the sandwich by turning both cams 180 so that the arrow points down Fig 4 10 x 10 5 cm gel plates clamped properly for gel casting If the plates are longer than 8 cm use spring clamps to secure the top section of the plates The notched plate faces the back block
8. to return the unit Pressure Casting clamp bar assembly 2 Silicone rubber gaskets 2 on each side 1 white under gasket and 1 black over gasket Casting cradle Cams 4 Pull cams out to disassemble the caster SA Fig 2 Location of T shaped spacers on notched plate Ni Fig 3 Align bottom edges of plates and spacers carefully Prepare the gel caster Oo Disassemble the caster Pull out both pairs of black cams at the side of the casting cradle and lift out both casting clamp assemblies Remove the black silicone rubber gasket from the bottom of the cradle and also the white foam gasket underneath it 124 Wash all gel caster components glass and alumina plates and spacers vvith a mild detergent Rinse thoroughly vvith deionized vvater Construct the gel sandyvich stack and place it into the caster Oo Loosen all 6 screws on the casting clamp assembly and lay it upright on a flat surface Slide both plastic pressure bars away from the back block Construct each gel sandwich For each sandwich choose one notched alumina or glass plate one rectangular glass plate and two spacers Use only unchipped plates Lay the notched plate on a flat surface place one spacer along each edge so that it aligns with the notch Fit a glass plate onto the spacers as shown The long flat side of the T shaped spacers fits between both plates The top of the T rests along the sides of the p
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