Mini-Blotting System, 4-place EBU-204 INSTRUCTION MANUAL
T A B L E O F C O N T E N T S Important User Information. . . . . . . . . . . . . . . . . . . . . . . . 3-4 Section 1: General Information 1.1 Introduction and Features . . . . . . . . . . . . . . . . . . . . . . . 5 1.2 Specifications and Constructions. . . . . . . . . . . . . . . . . . . . . 5 1.3 Safety. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
ENGLISH IMPORTANT USER INFORMATION This Instruction Manual will explain how to use this product safely and effectively. Please read and carefully follow the instruction manual in its entirety. The triangle/exclamation mark symbol alerts the user of the product to important operational, maintenance, and/or warranty requirements. The triangle/lighting bolt symbol alerts the user of the product to potentially hazardous electrical exposure.
FRANÇAIS INFORMATION IMPORTANTE À L’USAGE DES UTILISATEURS ESPAÑOL Le présent manuel d’utilisation explique la manière de se servir efficacement du produit en conditions de sécurité. Il est recommandé de soigneusement lire la totalité du manuel, avec ses consignes et ses instructions. El presente instructivo explica la manera de usar este producto en forma segura y efectiva. Sírvase leerlo en su totalidad y seguir detenidamente las indicaciones que contiene.
SECTION 1 General Information 1.1 Introduction and Features Electroblotting is a faster, easier, and more efficient method to permanently transfer PAGE gels to nitrocellulose and nucleic acids to nylon, than capillary (passive diffusion) blotting. For a similar transfer, traditional “capillary blotting” can take 6-24 hours to complete compared to only 45 to 90 minutes or less using electroblotting. The C.B.S.
1.4 Safety Power to the Electro-Blotter is to be supplied by an external DC voltage power supply that must be ground isolated so that the DC voltage output floats with respect to ground.
SECTION 2 Description of Parts 1.1 Unpacking the EBU-204 Please verify that your unit comes complete with the following components: · Blotting chamber with cooling base · Four Blotting cassettes with sponge pads · Safety cover with attached power leads · 2 electrode panels · Tubing adapters for cooling base 2.2 Components/Assembly 7 www.cbsscientific.
SECTION 3 Instructions for Use 3.1 Blotting Unit Preparation 1. 2. Place the blotting chamber on a level work surface in an authorized work area. Consult applications table 4.1 for type of transfer, buffer system, membrane type and power settings. Coolant Circulation: For coolant circulation, connect a heavy wall tubing which will not kink (hose clamps are optional), to the tubing adapters supplied with the unit. Excessive pressure will damage the plexiglass in the base and create leaks.
Figure 1 Figure 2 5. Assemble blotting stack as shown in Figure 3. With cassette wide open assemble components on black side in the following order: buffer saturated sponge pad, gel equilibrated in transfer buffer, buffer saturated transfer membrane, then buffer saturated blotting paper. Smooth with gloved finger or roll with glass rod to be sure no bubbles exist between the gel and the transfer membrane.
3.3 Standard Electro-Blot Transfer 1. Fill the chamber with transfer buffer. (Buffers should be prepared fresh with reagent grade chemicals and pre-cooled). The transfer buffer should come up to the top of the platinum labyrinth. The buffer should not come in contact with the banana plugs when the gel cassette sandwich(es) are immersed in the unit. 2. Align safety cover over the unit and carefully attach. Begin cycling of coolant. 3.
SECTION 4 Applications & Running Conditions for Tank Type Electro-Blotting 4.1 Recommended Buffers, Membranes, Power Settings and Transfer Times Gel Type Blotting Buffer Transfer Membrane Power denaturing: SDSPAGE (Western) ClearPAGE Transfer Buffer cat. # FB82500 as follows: for 1:20 dilution use 50ml ClearPAGE Transfer buffer, 200 ml Methanol and 770ml Ultrapure water nitrocellulose/ nylon PVDF: ionexchange · · · room temperature, 70V-100V, 200mA, 1.
4.2 References 1. 2. 3. 4. 5. 6. 7. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (ed.) (1993). Current Protocols in Molecular Biology. Vol. 2, Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., Ch.10. Burnette, W.N. (1981). Western blotting: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. Biochem.
SECTION 5 Maintenance of Equipment 5.1 Care and Handling The plastic components of the Electrophoretic Blotting units are fabricated from cell cast acrylic and polycarbonate. Electrodes and connectors are made from pure platinum, stainless steel, and chrome plated brass. As with any laboratory instrument, adequate care ensures consistent and reliable performance. After each use, rinse buffer chamber, cassettes and sponge pads with de-ionized water.
5.2 Maintenance The following inspection and maintenance procedures will help maintain the safety and reliable performance of the Blotting systems. Replacement parts can be ordered by calling 1-858-755-4959 or by contacting your local distributor. · · · Banana plugs and power cords should be inspected regularly. If the banana plugs become loose or do not feel friction tight replace the plugs or power cords. Should power cord assemblies (connectors, wire or shrouds) show any signs of wear or damage (e.
SECTION 6 Equipment and Accessories Cat. # Item EBU-204 Mini-Electrophoretic Blotting system, 4 gel cassette capacity. Includes electrophoresis chamber with cooling base and four 10 x 10cm cassettes. Removable platinum electrode panels, safety cover and interlocking power leads. EBX-BC-700 EBX-SP-700 EBX-EA-700 EBX-EC-700 Accessory Blotting Cassette, includes foam pad Accessory foam pads, set of 4 Replacement Anode Replacement Cathode POWER SUPPLIES Cat.
CONTACT INFORMATION Online Ordering www.cbsscientific.com Telephone Local and International: 858-755-4959 Toll Free: 800-243-4959 Sales E-mail Address sales@cbssci.com Technical Service E-mail Address technicalservice@cbssci.com Mailing Address P.O.
