Immobilon® PVDF membranes for Western blotting

How do Immobilon® membranes work?

Polyvinylidene fluoride (PVDF) membranes bind biomolecules through hydrophobic interactions. Membrane pores increase the surface binding area while restricting sizes of bound proteins.
 

We offer four different Immobilon® PVDF (Polyvinylidene Difluoride) membranes, each optimized for different protein blotting applications. We also now offer blotting sandwiches that feature pre-cut sheets of membrane and blotting filter paper.

  • Immobilon®-E membrane is the only PVDF membrane (0.45 µm) that wets out in aqueous buffers, eliminating the methanol pre-wet step. This membrane is well-suited for most Western blotting applications.
  • Immobilon®-P membranes are optimized for Western blotting and offer better handling and staining than nitrocellulose. All Immobilon-P PVDF membranes have a pore size of 0.45 μm.
  • Immobilon®-PSQ membrane (0.2 µm) is ideal for protein sequencing and immunoblotting of low molecular weight proteins. It has a higher protein binding capacity and a higher retention than 0.45 µm membranes.
  • Immobilon®-FL membrane (0.45 µm) was developed for fluorescence-based immunodetection. It has very low background fluorescence across a wide range of excitation and emission wavelengths.
  • Immobilon® NOW membrane offer the convenience of pre‑cut sheets with the flexibility of rolls in all formats: Immobilon®-E, Immobilon®-P, Immobilon®-FL and Immobilon®-PSQ membranes.

Immobilon® PVDF Membrane Advantages Compared to Nitrocellulose

  • Won’t crack or curl, and they can be cut without fracturing.
  • Low background
  • Broad solvent compatibility
  • Superior staining capabilities
  • Can be reprobed multiple times
  • Compatible with a variety of detection chemistries, including radioactive, chromogenic, chemiluminescent, fluorescent, and chemifluorescent techniques

Immobilon® PVDF membranes offer excellent protein binding and high signal.

Immobilon® PVDF membranes have high protein adsorption, so you won’t lose proteins during transfer or reprobing. The open pore structure makes it easy to access bound proteins and remove unbound probes. In addition, Immobilon® PVDF membranes optimized for fluorescent blots dramatically increase signal for high sensitivity in quantitative, multiplexing applications.
 

PVDF Membranes
Membrane
Name
Immobilon®-E membrane Immobilon®-P membrane Immobilon®-FL membrane Immobilon®-PSQ membrane
Pore Size 0.45 µm  0.45 µm 0.45 µm  0.2 µm
Detection
methods
Chemiluminescence, Colorimetric Chemiluminescence, Colorimetric Chemililuminescence, Colorimetric, Fluorescence (including near-infrared) Chemiluminescence, Colorimetric
Binding capacity Goat IgG 225-293 µg/cm2 294 µg/cm2 300 µg/cm2 448 µg/cm2
BSA n/a 215 µg/cm2 205 µg/cm2 340 µg/cm2
Insulin n/a 160 µg/cm2 155 µg/cm2 262 µg/cm2
Applications Western blots Western blots, dot blots Western blots, dot blots Western blots, dot blots
The durability and high protein retention of Immobilon® PVDF membranes make them
the preferred option for sequential detection of multiple targets via antibody stripping
and reprobing.
Selection
Criteria
• Target protein expressed over a broad dynamic range
• Wide range of molecular weights
• Target protein
<20 kDa
• Very low abundance target protein
• Low affinity antibody