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Immobilon Transfer Membranes, Sandwiches, and Blotting Filter Paper

Millipore offers three different Immobilon PVDF membranes, each optimized for different protein blotting applications. See below for available pre-cut sheet sizes and blotting filter paper.
 

Millipore offers three different Immobilon PVDF 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 membranes provide a number of advantages compared to nitrocellulose. They won’t crack or curl, and they can be cut without fracturing. They also have low background, broad solvent compatibility, and superior staining capabilities. In addition, they can be reprobed multiple times.

Immobilon-P Membrane

  • 0.45 µm pore size
  • Recommended for most western blotting, especially proteins >20 kDa
  • Millipore's Rapid Immunodetection Protocol reduces detection times by up to 2 hours by eliminating membrane blocking and several wash steps with no loss of sensitivity or specificity

Blotting Sandwiches

  • When you need to process multiple blots, blotting sandwiches are a convenient, time-saving choice. Our sandwiches include sheets of Immobilon-P membrane interleaved with pre-cut sheets of chromatography-grade blotting filter paper.

Blotting Filter Paper

  • Chromatography-grade blotting filter paper pre-cut to the most popular western blotting sizes.

Immobilon-PSQ Membrane

  • 0.2 µm pore size and a large internal structure
  • Higher protein adsorption and sequencing yields than other
  • membranes
  • Recommended for blotting proteins <20 kDa
  • Prevents blow-through of low molecular weight proteins

Immobilon-FL Membrane

  • The first transfer membrane optimized for fluorescence applications
  • Extremely low background improves sensitivity of all fluorescence detection protocols
  • Compatible with all commonly used fluorescent probes at all excitation and emission wavelengths
  • Ideal for multiplexing and chemifluorescence applications

Performance

Rapid Immunodetection Protocol with Immobilon-P Membrane

Step Standard Immunodetection Rapid Immunodetection
1. Block the membrane 1 h None
2. Incubate with primary antibody 1 h 1 h
3. Wash the membrane 3 x 10 min 3 x 5 min
4. Incubate with secondary antibody 1 h 30 min
5. Wash the membrane 3 x 10 min 3 x 5 min
6. Add substrate 5 min 5 min
Total time 4 h 5 min 2 h 5 min

Immobilon-PSQ Membrane Prevents Blow-through

Molecular 
weight standards (lanes 1, 3, 5, 7) and calf liver lysate (lanes 2, 4, 6, 8) 
were transferred to Immobilon-P or Immobilon-P<sup>SQ</sup> membranes. A sheet 
of Immobilon-P<sup>SQ</sup> was placed behind the primary membranes to capture 
proteins that passed through (lanes 5 and 6 behind Immobilon-P; lanes 7 and 8 
behind Immobilon-P<sup>SQ</sup>).

Molecular weight standards (lanes 1, 3, 5, 7) and calf liver lysate (lanes 2, 4, 6, 8) were transferred to Immobilon-P or Immobilon-PSQ membranes. A sheet of Immobilon-PSQ was placed behind the primary membranes to capture proteins that passed through (lanes 5 and 6 behind Immobilon-P; lanes 7 and 8 behind Immobilon-PSQ).

Immobilon-FL Membrane is Optimized for Fluorescence

Actin-tubulin multiplex detection on Immobilon-FL membrane. Rabbit muscle 
actin (red) was detected using rabbit anti-actin 1<sup>o</sup>AB and QDot® 655 
goat anti-rabbit 2<sup>o</sup>AB. Porcine brain tubulin (green) was detected 
using mouse anti-tubulin 1<sup>o</sup>AB and QDot 565 goat anti-mouse 
2<sup>o</sup>AB. Sensitivities down to 7 ng were observed for both analytes. 
Data provided by Quantum Dot Corporation.

Actin-tubulin multiplex detection on Immobilon-FL membrane. Rabbit muscle actin (red) was detected using rabbit anti-actin 1oAB and QDot® 655 goat anti-rabbit 2oAB. Porcine brain tubulin (green) was detected using mouse anti-tubulin 1oAB and QDot 565 goat anti-mouse 2oAB. Sensitivities down to 7 ng were observed for both analytes. Data provided by Quantum Dot Corporation.


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