INSTRUCTION MANUAL Electrophoretic Blotting System EBU-222 www.cbsscientific.
TABLE OF CONTENTS Page Important User Information 3-4 Section 1 1.1 1.2 1.3 General Information Introduction 5 Specifications 5 Safety 5 Section 2 2.1 2.2 Description of parts Unpacking EBU-222 6 Components/Assembly 6 Section 3 3.1 3.2 3.3 3.4 Instructions for Use Blotting Unit Preparation 7 Preparing the Gel for Transfer 7-8 Standard Electro-Blot Transfer 8 Removing the Cassettes 8 Section 4 4.1 4.
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 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. Le triangle avec point d’exclamation est un symbole destiné à avertir l’utilisateur du produit de l’importance de certaines exigences relatives au fonctionnement, à l’entretien et/ou à la garantie.
SECTION 1 General Information 1.1 Introduction 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 2 hours or less using electroblotting. The C.B.S.
SECTION 2 DESCRIPTION OF PARTS 2.1 • • • • • Unpacking the EBU-222 Please verify that your unit comes complete with the following components: Blotting chamber Blotting cassette with sponge pads Safety cover with attached power leads 2 electrode panels Tubing adapters for cooling base 2.2 Components/Assembly To open gel cassette lift white latch as shown by green arrow. Cassette includes 2 sponge pads.
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.
CASSETTE ASSEMBLY 9. Remove entire assembly from pyrex dish and load into desired position (slot) in buffer chamber with black screen facing towards black terminal (cathode) and red screen facing towards red terminal (anode). 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.
SECTION 4 APPLICATIONS & RUNNING CONDITIONS FOR TANK TYPE ELECTRO-BLOTTING 4.1 RECOMMENDED BUFFERS, MEMBRANES, POWER SETTINGS AND TRANSFER TIMES Blotting Buffer Transfer Membrane denaturing: SDS-PAGE (Western) Gel Type Towbin (25mM Tris/192mM Glycine/ pH 8.3/ Methanol 20% (w/v) nitrocellulose/nylon PVDF: ion-exchange • room temperature, 70V-100V, 200mA, 1.5- 4 hours • 4ºC, 150V, 400mA, .
SECTION 5 MAINTENANCE OF EQUIPMENT 5.1 CARE AND HANDLING The plastic components of the Electrophoretic Blotting units are fabricated from 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 Scotch Brite pads with de-ionized water.
SECTION 6 EQUIPMENT AND ACCESSORIES Cat. # Item EBU-222 EBE-202 EBE-202 EBC-202 EBS-202 Electrophoretic Blotting System. Includes chamber with 2 platinum wire electrode panels, 1 colorcoded gel cassette, safety cover with power leads electrode panels, safety cover and interlocking power leads. Replacement Electrode Panel for EBU-222 Replacement Electrode Panel for EBU-222 Additional Gel Cassette, color coded Sponge Pads POWER SUPPLIES Cat. # Item EPS-200 X EPS-600 EPS-300-II EPS-300-IIV www.
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