Thermo Scientific OWL HEP-1 Operating instructions

Semidry Electroblotter
Models HEP-1 and HEP-3
Operating and Maintenance Manual 7007332 Rev. 0
Visit us online to register your warranty
www.thermoscientific.com/warranty

Thermo Scientific
MANUAL NUMBER 7007332
0 -- 5/1/12 Transfer to Marietta (was The Panther 3/2003) ccs
REV ECR/ECN DATE DESCRIPTION By
Preface
Semidry Electroblotter i

Thermo Scientificii Semidry Electroblotter
Preface
Important Read this instruction manual. Failure to read, understand and follow the instructions in this manual
may result in damage to the unit, injury to operating personnel, and poor equipment performance. s
Caution All internal adjustments and maintenance must be performed by qualified service personnel. s
Warning To avoid the risk of personal shock, always disconnect the gel box from the power supply. Further,
the power supply must be equipped with a shut-down-on-disconnect circuit. Running conditions for this unit
should not exceed the name plate readings found on the lower buffer chamber. s
This system is designed to meet IEC 1010-1 safety standards (IEC 1010-1 is an internationally accepted
electrical standard for laboratory instruments).
Statement of Proper Use: Use this product only for its intended purpose as described in this manual. Do not
use this product if the power leads are damaged or if any of its surfaces are cracked.
Material in this manual is for information purposes only. The contents and the product it describes are subject
to change without notice. Thermo Fisher Scientific makes no representations or warranties with respect to this
manual. In no event shall Thermo Fisher Scientific be held liable for any damages, direct or incidental, arising
out of or related to the use of this manual.
©2012 Thermo Fisher Scientific. All rights reserved.

Thermo Scientific Semidry Electroblotter iii
Preface
Important operating and/or maintenance instructions. Read the accompanying text carefully.
Potential electrical hazards. Only qualified persons should perform procedures associated with this
symbol.
Equipment being maintained or serviced must be turned off and locked off to prevent possible injury.
Hot surface(s) present which may cause burns to unprotected skin, or to materials which may be
damaged by elevated temperatures.
Marking of electrical and electronic equipment, which applies to electrical and electronic equipment
falling under the Directive 2002/96/EC (WEEE) and the equipment that has been put on the market
after 13 August 2005.
This product is required to comply with the European Union’s Waste Electrical & Electronic
Equipment (WEEE) Directive 2002/96/EC. It is marked with the WEEE symbol. Thermo Fisher
Scientific has contracted with one or more recycling/disposal companies in each EU Member State
European Country, and this product should be disposed of or recycled through them. Further
information on Thermo Fisher Scientific’s compliance with this directive, the recyclers in your country
and information on these products will be available at www.thermofisher.com.
4Always use the proper protective equipment (clothing, gloves, goggles, etc.)
4Always dissipate extreme cold or heat and wear protective clothing.
4Always follow good hygiene practices.
4Each individual is responsible for his or her own safety.

Thermo Scientificiv Semidry Electroblotter
Preface
Do You Need Information or Assistance on
Thermo Scientific Products?
If you do, please contact us 8:00 a.m. to 6:00 p.m. (Eastern Time) at:
1-740-373-4763 Direct
1-800-438-4851 Toll Free, U.S. and Canada
1-877-213-8051 FAX
http://www.thermoscientific.com Internet Worldwide Web Home Page
[e-mail] Tech Support Email Address
Certified Service Web Page
Thermo Fisher Scientific
401 Millcreek Road, Box 649
Marietta, OH 45750
Our staff can provide information on pricing and give you quotations. We can
take your order and provide delivery information on major equipment items or make
arrangements to have your local sales representative contact you. Our products are listed on the
Internet and we can be contacted through our Internet home page.
Our staff can supply technical information about proper setup, operation or
troubleshooting of your equipment. We can fill your needs for spare or replacement parts or
provide you with on-site service. We can also provide you with a quotation on our Extended
Warranty for your Thermo Scientific products.
Whatever Thermo Scientific products you need or use, we will be happy to discuss your
applications. If you are experiencing technical problems, working together, we will help you
locate the problem and, chances are, correct it yourself...over the telephone without a service
call.
When more extensive service is necessary, we will assist you with direct factory trained
technicians or a qualified service organization for on-the-spot repair. If your service need is
covered by the warranty, we will arrange for the unit to be repaired at our expense and to your
satisfaction.
Regardless of your needs, our professional telephone technicians are available to assist you
Monday through Friday from 8:00 a.m. to 6:00 p.m. Eastern Time. Please contact us by
telephone or fax. If you wish to write, our mailing address is:
International customers, please contact your local Thermo Scientific distributor.
Sales Support
Service Support
www.unitylabservices.com

Semidry Electroblotter vThermo Scientific
Table of Contents
General Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-1
Setting Up . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-1
Materials Needed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-1
Setting Up the Blot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-2
DNA Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-3
Gel Sandwich Assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-4
Using the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-1
Running the Blot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-1
Transfer settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-1
Running Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-2
Factors That Affect Transfer Efficiency . . . . . . . . . . . . . . . . . . . . . .3-2
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-1
Technical Tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-1
Semidry Blotting - General References . . . . . . . . . . . . . . . . . . . . . .5-2
Recipes for Buffers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-3
Care and Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-1
Optional Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-1
Section 1
Section 2
Section 3
Section 4
Section 5
Section 6
Section 7

Semidry Electroblotter 1-1Thermo Scientific
Section 1 General Information
The Thermo Scientific semi-dry electroblotter offers rapid transfer of
proteins or nucleic acid molecules from polyacrylamide or agarose gels to
membranes. Solid plate platinum/titanium and stainless steel electrodes are
highly conductive and allow transfer at low voltages without external
cooling systems. Plate electrodes also provide a uniform electric field for
efficient, even transfers.
Before starting, unpack the unit and inventory your order. If any parts are
missing, contact Technical Services immediately. You have 30 days from
date of shipment to make any shipment claims.
Item Description . . . . . . . . . . . . . . . . . .Catalog No.
Complete System . . . . . . . . . . . . . . . . . . . . . .HEP-1
Transfer Area . . . . . . . . . . . . . . . .20cm W x 20cm L
Footprint . . . . . . . . . . .25cm W x 25cm D x 5cm H
Complete System . . . . . . . . . . . . . . . . . . . . . .HEP-3
Transfer Area . . . . . . . . . . . . . . . .35cm W x 45cm L
Footprint . . . . . . . . . . .40cm W x 51cm D x 5cm H

1-2 Semidry Electroblotter Thermo Scientific
Section 1
General Information
Heavy Duty Knobs
HEP-1 (3)
HEP-3 (4)
Power Supply Leads
Lid
Anode
Cathode
Base
Figure 1-1. Exploded View
Table 1-1. Parts List
• 1 base with stainless steel cathode
• 1 lid with platinum/titanium anode
• 1 attached pair of power supply leads
• 3 knobs for the HEP-1, 4 knobs for the HEP-3

Semidry Electroblotter 2-1Thermo Scientific
Section 2 Setting Up
Once the proteins (or nucleic acids) in a sample aliquot have been
separated on a slab gel, the resulting bands may be transferred to a solid
support membrane. The primary reason for this type of blot is one of
localization and secondarily, concentration of discrete bands. Although
many have used alternative cross linking agents such as DATD
(N,N'dihydroxyethylene-bis Acrylamide) to allow for the accessibility of
gel bound proteins, this still represents an impediment to radio
enumeration due to the quenching by the gel matrix itself.
The most common solid support membrane is nitrocellulose. A second
type of membrane is PVDP (Polyvinylidene difloride) which is generally
used when a transferred protein is to be sequenced, additionally, it has a 2x
binding capacity. Also used for nucleic acid capture are Nylon membranes.
In either case, the proteins are transferred from the gel to the matrix in an
electric field perpendicular to the gel (initial running direction).
Tris based buffers are employed in the transfer. Methanol and SDS are
modifiers often use in protein transfer buffer. These components however
are antithetical in their effects both in terms of movement and adsorption.
Methanol restricts protein movement from the gel but is often required to
support the ionic nature of protein to nitrocellulose binding. SDS aids in
protein elution, but can also inhibit binding of small molecular weight
proteins (Mozdzanowski, J., High yield electroblotting, Electrophoresis,
1992, Vol l3.,p.59-64).
In order to use this blotting device, you will need:
Power supply - Blotting requires a power supply that can operate at a fairly
high current setting and low voltage. If an inappropriate power supply is
used, the power supply may blow a fuse, shut itself off, display a no load or
short load message or even have a short circuit. It is very important to be
sure that the power supply you will be using will work with this device.
Some power supplies that will work with this device are Catalog No. OSP-
135, OSP-300, EC Apparatus EC135 and EC570, and Bio Rad's
PowerPac 200. If your power supply is not among those listed, contact the
manufacturer of your power supply to determine if it will work for tank
blotting applications. Contact Technical Services for power supply
recommendations if you do not have an appropriate power supply.
Materials Needed