Related Manuals for Thermo Scientific OWL HEP-1
Summary of Contents for Thermo Scientific OWL HEP-1
- Page 1 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...
- Page 2 Preface MANUAL NUMBER 7007332 5/1/12 Transfer to Marietta (was The Panther 3/2003) ECR/ECN DATE DESCRIPTION Thermo Scientific Semidry Electroblotter...
- Page 3 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. Semidry Electroblotter Thermo Scientific...
- Page 4 Always use the proper protective equipment (clothing, gloves, goggles, etc.) 4 Always dissipate extreme cold or heat and wear protective clothing. 4 Always follow good hygiene practices. 4 Each individual is responsible for his or her own safety. Thermo Scientific Semidry Electroblotter...
- Page 5 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...
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Page 6: Table Of Contents
Optional Equipment ......... . .7-1 Thermo Scientific... -
Page 8: General Information
General Information Section 1 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. -
Page 9: Section 1 General Information
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 Thermo Scientific... -
Page 10: Setting Up
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. Thermo Scientific Semidry Electroblotter... -
Page 11: Setting Up The Blot
Incubate the gel in transfer buffer for 15 min. with gentle agitation. If the gel is on the plate, it will become loose during this step. Semidry Electroblotter Thermo Scientific... -
Page 12: Dna Gels
A small portion of the gel may still be attached to the glass; in this case, a small stream of water from a wash bottle can be used to aid in removal of the gel. Thermo Scientific Semidry Electroblotter... -
Page 13: Gel Sandwich Assembly
7. Wearing gloves, gently smooth out any bubbles by forcing them to the closest edge of the gel. Test tubes and pipettes many also be used for this purpose. Semidry Electroblotter Thermo Scientific... - Page 14 10. Repeat with three more pieces of filter paper. 11. Holding the stack, drain off all excess buffer from the plate. Wipe away any droplets around the edge of the stack. Anode Filter Paper Transfer Membrane Filter Paper Cathode Figure 2-1. Assembly Order Thermo Scientific Semidry Electroblotter...
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Page 16: Using The System
Blots may also be run at constant voltage. Some power supplies have difficulty sustaining steady voltages at these low voltage settings. If you find that voltages are fluctuating, or that the power supply shuts itself off when set on constant voltage, use constant current settings instead. Thermo Scientific Semidry Electroblotter... -
Page 17: Running Conditions
~5-8V 30 minutes to 2 hours Needs to be experi- 30 minutes to 2 hours, 15-20 minutes at this Running Time mentally determined generally in the lower setting (large molecules need range longer transfer times) Semidry Electroblotter Thermo Scientific... -
Page 18: Troubleshooting
• Filter paper should be saturated with transfer • The filter paper was too dry buffer before adding them to the sandwich. Thermo Scientific Semidry Electroblotter... -
Page 19: Section 4 Troubleshooting
• Too much current high in ionic strength can also burn a gel by over- heating. A cracked and dry gel often is an indica- tor of overheating. Semidry Electroblotter Thermo Scientific... -
Page 20: Technical Tips
"hybridized" or attached to specific pieces of DNA. Northern Blotting is a similar method but the molecules involved are RNA. Both Southern and Northern blotting generally require the DNA or RNA to first be separated out on an agarose gel. Thermo Scientific Semidry Electroblotter... -
Page 21: Semidry Blotting - General References
7. Dunbar, B.S., Ed. 1994. Protein Blotting: A Practical Approach. IRL Press at Oxford University Press, Oxford, England. A great guide to blotting techniques, including visualization, immunological techniques, and sequence analysis. Semidry Electroblotter Thermo Scientific... -
Page 22: Recipes For Buffers
1X Tris-glycine buffer (Towbin buffer minus the methanol) is used for agarose and polyacrylamide gel electrophoresis of nucleic acids. Towbin buffer (containing 20% methanol) is a commonly used buffer for semi- dry transfers. 0.025M Tris Base 0.192M Glycine 20% MeOH pH 8.3 Thermo Scientific Semidry Electroblotter... - Page 23 For transfer of RNA from agarose gels. With its high buffering capacity and low ionic strength, this buffer is more efficient than TAE, TBE or MOPS from agarose gels. 50X: 0.2M morpholinopropanesulfonic acid (MOPS) 50mM sodium acetate 5mM EDTA pH 7.0 Semidry Electroblotter Thermo Scientific...
- Page 24 0.8M Tris 1.18M borate 24mM EDTA pH 8.3 CAPS Buffer pH 11 This buffer can be used to improve transfer of some proteins. 10mm CAPS (3-[cyclohexylamino]-1-1 0% methanol propanesulfuric acid, adjust to pH 11 with sodium hydroxide. Thermo Scientific Semidry Electroblotter...
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Page 26: Care And Cleaning
To order RNase Away®, contact Technical Services: P/N 21-236-21 250ml bottle, P/N 21-402-178 475ml spray bottle, P/N 14-375-35 1 liter bottle, P/N 14-754-34 4 liter bottle *Rnase AWAY® is a registered trademark of Molecular BioProducts, Inc. Thermo Scientific Semidry Electroblotter... -
Page 27: Section 6 Care And Cleaning
This list does not include all possible chemical incompatibilities and safe compounds. Acrylic products should be cleaned with warm water, a mild detergent such as Alconox™, and can also be exposed to a mild bleach solution (10:1). In addition, RNAse removal products are also safe for acrylic. Semidry Electroblotter Thermo Scientific... -
Page 28: Optional Equipment
ER-35 Electroblot Buffer Kit For optimal performance on Western blotting applications, the ER-35 electroblot buffer kit is recommended. These three different buffers (anode 1, anode 2 and cathode 1) vary in buffering capacity and increase the transfer efficiency. Thermo Scientific Semidry Electroblotter... - Page 30 Thermo Scientific Semidry Electroblotter...
- Page 31 Section 8 Warranty Information Semidry Electroblotter Fisher Scientific...
- Page 32 Thermo Fisher Scientific 401 Millcreek Road Marietta, Ohio 45750 United States www.thermofisher.com...
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