Hoefer SE640 Manuel d'utilisateur

user manualum    SE640-IM/Rev.B0/07-12Hoefer SE640Wide-mini Dual Gel Electrophoresis Unit

• p2Specifications Gel plate size (w × h): 18 × 8 cmGel size: 14 × 8 cmMaximum watt: 50 WMaximum volt: 1000 VMaximum ampere: 500 mAMaximum tempera

• p3Fig 1. Main components of the SE640 series (see Fig 4 for caster components).Included but not shown:• Gel Seal compound, ¼ oz. • Spacer-Mate spa

• p4Unpacking and inventoryUnwrap all packages carefully and compare con-tents with the packing list, making sure all items arrived. If any part is mi

• p5Glass  platesThe plates are 18 cm wide and 8 cm in length Three sets of glass plates are included with each unit. Notched divider plates, ordered

• p6Spacers(May be ordered separately.) Spacers determine the thickness of the gel and are available in three thicknesses (0.75, 1.0, and 1.5 mm).Spac

• p7Operating instructionsGel casting and electrophoresis procedures follow. Included are instructions for polyacryl-amide gels (used with continuous

• p8Construct the  gel sandwich and insert  into caster1Prepare the caster and clampsPlace the spirit level into the caster center and adjust the leve

• p9Club  sandwichA notched center divider plate (ordered separately) pairs two sandwiches to double the number of gels that can be cast and run.Assem

• p105Place the laminated gasket into the casting cradle (See Fig 4) with the foam side down. Place the clamp assembly in the casting cradle, screw si

• p11Acrylamide gels1Prepare the monomer solution and pour the gelSee Appendix A for SDS-PAGE recipes. Prepare the required amount of monomer solution

• piContents Important Information ... iiWaste Electrical and Electronic Equipment (WEEE) ...

• p122Overlay each gel with a thin layer of water-saturated n-butanol, water, or diluted gel buffer to prevent gel exposure to oxygen. Slowly deliver

• p13Gradient gelsBoth linear and exponential gradient gels can be poured in the dual gel caster. We recommend using a Hoefer SG Series Gradient Maker

• p144Pour the “light” solution into the reservoir chamber (the chamber furthest from the inlet). Open the stopcock long enough to displace air betwee

• p15Sample preparation  and loadingThe sample can be loaded either while the sand-wich is in the caster or after the upper buffer chamber is attached

• p163Heat the tube in boiling water for 90 seconds, then allow to cool to room temperature. Treated samples can be stored at -40 to -80 °C for future

• p17Final assembly Upper buffer  chamber1Rinse both buffer chambers with water and distilled water thoroughly before each use. Clean away any gel adh

• p184Use a pipet to carefully fill each slot above the sample wells with buffer in order to minimize disturbing the samples. Then pour 100 ml of buffe

• p195Plug the color-coded leads into the jacks of an approved power supply. Plug the red lead into the red output jack and the black lead into the bl

• p20Separating the sampleElectrophoresis  parameters for discontinuous  polyacrylamide  gelsGels may be run at either constant current or constant vo

• p21TimeA run is complete when the tracking dye reaches the bottom of the gel. A 1.5-mm thick Laemmli SDS gel, run at 25 mA/gel without cooling, usu-

• piiImportant Information  – English• If this equipment is used in a manner not speci-ed by Hoefer, Inc. the protection provided by the equipment m

• p22After electrophoresis1Once the tracking dye reaches the bottom of the gel, turn off the power supply, disconnect the leads and remove the safety

• p23Care and maintenanceCleaningImmediately after each use, rinse the upper and lower buffer chambers with water and then rinse thoroughly with disti

• p24Customer service informationTechnical service and repairHoefer, Inc. offers complete technical support for all of our products. If you have any q

• p25Troubleshootingproblem possible cause remedy Gel sandwich  leaks while  castingDirty or damaged components Plates, spacers, and the gasket must

• p26Upper buffer  chamber leaksMis-aligned parts Check that the glass plates, spacers, and clamps are aligned and fit snugly into the upper chamber g

• p27Bands are  skewed or distortedIncomplete gel preparation and polymerizationDegas the stacking-gel solution and avoid trapping air bub-bles under

• p28Poor band  resolutionRunning conditions Begin electrophoresis as soon as the sample is loaded to pre-vent low molecular weight species from diff

• p29Appendix A:  Laemmli system gelsThe Laemmli system is the most common elec-trophoresis protocol for SDS-denatured proteins. The leading ion in th

• p30The total percent of acrylamide (% T) in the resolving gel, which can range from 5 to 20%, determines the pore size. Commonly, the amount of cros

• p31Solutions 1.  Acrylamide stock solution (30.8% T 2.6% C Bis, 200 ml) Acrylamide (FW 71.08) 30% w/v 60 g Bis* (FW 154.2) 0.8% w/v 1.6 g Deion

• piiiBelangrijke Informatie  – Dutch• Indien deze uitrusting in een manier wordt gebruikt die niet door Hoefer, Inc. is gespeciceerd de bescherming

• p326.  0.375 M TrisCl, 0.1% SDS, pH 8.8 (Resolving gel overlay, 100 ml) 1.5 M Tris-Cl, pH 8.8 (Soln. #2) 0.375 M 25.0 ml 10% SDS (Soln. #4) 3.

• p338.    0.025 M Tris, 0.192 M glycine, 0.1% SDS, pH 8.3 (Electrophoresis buffer, 5.0 liters) Tris (FW 121.1) 0.025 M 15.1 g Glycine (FW 75.07)

• p34Gel recipes The Laemmli gel recipes are for 30 ml of a single concentration solution (enough for two 1.5 mm, 18 × 8 cm gels). Tabulated are ingre

• p35For linear gradient gels, use equal volumes of low % and high % acrylamide solutions in the 2 chambers of the gradient maker. Less APS is added t

• p36Appendix B: BibliographyGeneralGallagher, S. R., and Smith, J. A., Electrophoretic separation of proteins. In Current Protocols in Molecular Biol

• p37Two-dimensional electrophoresisAdams, L. D. and Gallagher, S. R., Two-Dimensional Gel Elec-trophoresis Using the O’Farrell System. Current Protoc

• p38Ordering informationproduct  quantity  code no.SE640 Dual Vertical Unit, basic. 1 SE640 Includes: 3 sets of glass plates, four 8 cm clamp

• p39basic caster SE6015cam SE6005Luniversal clamp SE6403U spirit level SER11gasket SE6009safety lid with cables SE6056upper buffer chamber with ele

• p40Combs number  thickness   width    of wells  (mm)  (mm)  quantity  code number 10 0.75 8.3 1 SE511-10-.75 10 1.00 8.3 1 SE511-10-1.0 1

• p41Spacers  thickness     width   (mm)  length  (mm)  quantity  code no. 0.75 8 2 2 SE6419-2-.75 1.00 8 2 2 SE6419-2-1.0 1.50 8 2 2 S

• pivdes dommages irréparables à l’unité!• Ne pas fonctionner avec les températures de tampon au-dessus du maximum a spécié des spécications techni

Hoefer, Inc. 84 October Hill Road Holliston, MA 01746Toll Free: 1-800-227-4750 Phone: 1-508-893-8999 Fax: 1-508-893-0176 E-mail: [email protected]

• pv• Sikkerheten lokket må være på plass før forbinding kraftforsyningene blyene til en kraftforsyning. • Vender all kraftforsyningsstyring av og f

• pvipor Hoefer, Inc. puede ser utilizado para operar, para mantener, y para atender a este producto.• Sólo utiliza una alimentación que es CE marcó

• pviiEnglishFrenchGermanItalianSpanishSwedishThis symbol indicates that the waste of electrical and elec-tronic equipment must not be disposed as uns

• p1 Gel electrophoresis unit  function and descriptionThe Hoefer SE600 series vertical slab gel electrophoresis units (SE600, SE640 and SE660) are i