Hoefer SE900 Manuel d'utilisateur

user manualum SE900-IM/Rev.B0/06-12Hoefer SE900Second Dimension Electrophoresis System

• p4Separation TankThe Separation tank is transparent to allow visualization of the track-ing dyes during electrophoresis. The chamber base contains a

• p5Safety Interlocks Safety interlocks are mounted on the sides of the tank. One is red and the other is black to help with orienting the gels correc

• p6PAGE RackThe rack serves three purposes:1. It divides the tank into anodic and cathodic chambers and supports and seals the gel cassettes vertica

• p7Safety LidThe safety lid holds the high voltage leads that connect to an external power supply (not supplied). The lid is held in place by the saf

• p8Multiple Gel Caster The multiple gel caster is included with the SE900-1.0 (Fig 10), and can be ordered separately as the SE915. The multiple gel

• p9Separator SheetsThe sheets have a protective film on both sides which should be removed before use.Wonder WedgeThe Wonder Wedge is helpful to pry a

• p10Operating InstructionsSeparation Tank Setup1 Position the SE900 with the black safety interlock on the left, as shown in Fig 20 on page 17. 2

• p11Using the Multiple Gel CasterCasting Homogenous Gels1 Make sure the caster and the glass cassettes are clean and dry.2 Close each of the glass

• p12Prepare the Acrylamide Solution1 See Appendix A for SDS-PAGE recipes. For casting six SE9102-1-1.0 gels, 400 ml is needed. 2 De-aerate the ge

• p13Gradient GelsLinear gradient gels can be cast using an optional accessory, the Hoefer SG500 Gradient Maker. The SG500 mixes low percentage and hi

• piPage Finder Important Information ...iiWaste Electrical And Electronic Equipment (WEE

• p14Disassembling the Multiple Gel Caster1 Place the caster down on its back in a tray or sink.2 Remove the red clamps and screws securing the fa

• p15Sealing the IPG to the Second Dimension Gel1 Remove residual liquid from the top of the second dimension gel by tipping the cassette and blotti

• p16Page Separation 1 Remove the safety lid. 2 Fill the tank with 12 liters of 1X electrophoresis buffer. This can be prepared in the separation

• p17Fig 20. Gel orientation.Fig 21. Slide safety lid into place.Note: The black high voltage lead should be on the same side as the IPG strip.Blac

• p18After Electrophoresis1 Unplug the leads to the power supply.2 Remove the lid by moving the safety interlocks while lifting the lid.3 Care

• p19Care and Maintenance• Handle the PAGE rack with care to prevent damage to the banana plugs and electrode wires. • Do not drop items onto the ce

• p20Troubleshooting Problem Solution Casting Issues: Gel caster leaks If the stack is too tall, the front plate may not seat firmly against the gas

• p21Problem Solution Gradient gels-uneven layering Add sucrose (15% final concentration) or glycerol (25% final concentration) to the high-percent m

• p22Problem Solution Instrument Problems: No voltage or current at start of the run Instrument not properly attached to power supply. Make sure hi

• p23Ordering Information Product Quantity Order CodeLarge Format PAGE 1 SE900 Large Format PAGE Complete 1 SE900-1.0Tubing Kit for Recircula

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

• p24Appendix A: Theory and RecipesLaemmli System GelsThe Laemmli system is the most common electrophoresis protocol for SDS-denatured proteins.The le

• p25Solutions1. Acrylamide Stock Solution(30.8% T 2.6% C Bis, 1000 ml)Acrylamide (FW 71.08) 30% w/v 300 gBis*N,N' Methylenebisacrylamide

• p266. 0.375 M Tris-Cl, 0.1% SDS, pH 8.8(Resolving gel overlay, 100 ml)1.5 M Tris-Cl, pH 8.8 (Solution #2) 0.375 M 25 ml10% SDS (Solution #4

• p279. 1% Agarose in 1X Electrophoresis Buffer (100 ml)In a 500 ml flask add:Agarose 1 g10X Electrophoresis Buffer (Solution #8) 10 mlDeionized

• p28Gel RecipesThe Laemmli gel recipes are for 400 ml of a single concentration solu-tion (enough for six 1.0 mm, 25 × 20 cm gels). Tabulated are vol

• p29Laemmli Gel (per 200 ml each gel solution)Gradient Gels Solution 200 ml LIGHT 5.0% 7.5% 10.0% 12.5% 15.0% 17.5%Acrylamide Stock (Soln. #1)

• p30Appendix B: ReferencesDenaturing Gel SystemsLaemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T. Na

Hoefer, Inc.84 October Hill Road Holliston, MA 01746 Toll Free: 1-800-227-4750 Phone: 1-508-893-8999 Fax: 1-508-429-5732E-mail: [email protected]

• piiikäyttöjännitelyijyjä käyttöjännitteeseen. • Kiertää kaikki käyttöjännitevalvonnat ja irrottaa valtalyijyt ennen poistaminen turvallisuuskantta.

• pivtrykket er unregulated. • Introduserer Aldri antifreeze eller noe organisk løsemiddel inn i noe del av instrumentet. Organiske løsemiddler vil f

• pvWaste Electrical And Electronic Equipment (WEEE) This symbol indicates that the waste of electrical and electronic equipment must not be disposed

• p1Function and DescriptionThe Hoefer® SE900 vertical slab gel electrophoresis unit is intended for the second dimension of 2D electrophoresis. It is

• p2Specifications Gel plate size (w × h): 28 × 21 cmGel size: 25 × 20 cmMaximum watt: 100 WMaximum volt: 600 VMaximum ampere: 1000 mAMaximu

• p3Unpacking and InventoryUnwrap all packages carefully and compare the contents with the packing list, making sure all the items have arrived. If an