Preparation of semi-dry transfer buffer

Preparation of Semi-dry Transfer Buffer

  1. Cut the transfer membrane and four pieces of Whatmann 3 MM paper to gel size (7 x 8 cm).

  2. Equilibrate the membrane in transfer buffer. By use of PVDF membranes equilibrate the membrane first for 2 minutes in methanol and then for additional 5 minutes in transfer buffer.

  3. Wet the porous pads as well as the four pieces of Whatmann 3 MM paper with transfer buffer.

  4. Remove the gel from the cassette and equilibrate the gel for 5 minutes in transfer buffer.

  5. Mount the transfer sandwich analogue to the tank blot sandwich and place it into the semi-dry blotter.

  6. Transfer is done at room temperature with 0.8 - 1.5 mA/cm2 gel area for approx. 1 hour. 


By blotting of proteins of differently large sizes the use of a discontinuous blotting buffer system is recommended.

 

Transfer buffer:

Anode buffer 1: 300 mM Tris, pH 9.5
Anode buffer 2: 30 mM Tris, pH 9.5
Cathode buffer: 40 mM 6-Aminohexanoic acid, 20 % (v/v) methanol

 

Blotting sandwich:

Anode (+)

  • 2 filter papers moisted with anode buffer 1
  • 2 filter papers moisted with anode buffer 2
  • Transfer membrane
  • 3 filter papers moisted with cathode buffer

Cathode (-)


Top
Close window
SERVA Market Square 2021: September - December

• Gel Media
• Protein & DNA Standards
• Protein Quantification
• Buffers & Buffer Solutions
• Detergents

 

→ This way to the Special Offer

Close window
Product of the Month September:
AquaSpark™ Substrate

AquaSpark™ Substrate for Chemiluminescence Detection of Alkaline Phosphatase (AP) in ELISA and Blot

→ This way to the Special Offer

SERVA InfoMail - Get the latest information

Close window

When subscribing to »SERVA InfoMail« you will receive information about the latest product release, special promotions, new jobs at SERVA and more. This service is, of course, free of charge.
You may remove your name any time from the list of recipients.

Quick Order

Close window

Using the Quick Order field, you can add a product to your shopping cart with just one click. All you have to do: enter the Cat.No. as shown in our catalog in the format xxxxx.yy and click the Go!-Button.