Consistent Western Blot Transfer with Wet Tank Systems
Автор: Thermo Fisher Life Sciences
Загружено: 2025-10-29
Просмотров: 27
Описание:
Learn a dependable wet tank western blot transfer workflow: https://www.thermofisher.com/us/en/ho...
What you’ll learn
When to choose wet transfer vs semi-dry
PVDF vs nitrocellulose and how to pre-wet/equilibrate
Stack order (cathode→gel→membrane→anode) & bubble removal
Current/time settings, agitation, and cooling/ice pack use
Quick post-transfer handling for best detection
Materials
Wet tank transfer unit + power supply
Transfer buffer (cold), ice/ice packs, stir bar or rocker
Polyacrylamide gel; PVDF or nitrocellulose membrane
Sponges/filter papers, roller/forceps, timer
Steps
1. Chill buffer and pre-wet membrane; equilibrate gel.
2. Assemble stack in cassette (ensure correct orientation).
3. Fill tank with cold buffer; insert cassettes; add cooling/ice pack.
4. Set current/time; start run with gentle agitation.
5. Remove membrane, mark lanes, proceed to blocking/detection.
FAQs:
1. Bands look “smiley”? Lower current, improve cooling, ensure even contact.
2. Weak transfer of large proteins? Increase time, include SDS per protocol, confirm PVDF use.
#WesternBlot #WetTransfer #ProteinTransfer
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