Nitrocellulose Membranes for Western Blotting
Nitrocellulose membranes are a popular matrix used in protein blotting because of their high protein-binding affinity, compatibility with a variety of detection methods (chemiluminescence, chromogenic, and fluorescence), and the ability to immobilize proteins, glycoproteins, or nucleic acids. Protein immobilization is thought to occur by hydrophobic interactions, and high salt and low methanol concentrations helpimprove protein immobilization to the membrane during electrophoretic transfer, especially for proteins with higher molecular weights. Nitrocellulose membranes are not optimal for electrophoretic transfer of nucleic acids, as the high salt concentrations that are required for efficient binding will effectively elute some or all of the charged nucleic acid fragments.
- High-quality—pure, 100% nitrocellulose membranes with high surface area and excellent uniformity
- Selection—available in 0.2 µm and 0.45 µm pore sizes, nitrocellulose varieties for peptide and protein applications, respectively
- Convenient—available as ready-to-use pre-assembled membrane/filter paper sandwiches or several sizes of pre-cut sheets or economically priced rolls for cutting to any dimension
- High binding affinity—provides high affinity protein binding, blocks easily, and exhibits very low background in chemiluminescent western blotting
Figure 1. Nitrocellulose blot of E-PAGE™ Gel developed with WesternBreeze® Kit using Invitrogen™ Nitrocellulose Membranes. MagicMark™ XP Western Protein Standard (5 µL) was loaded into each well of an E-PAGE™ 48 8% Gel and transferred to a pre-cut 0.2 µm nitrocellulose membrane. The blot was probed with 1:1000 Anti-Xpress™ Antibody and detected using the WesternBreeze® Chemiluminescent (Panel A) or WesternBreeze® Chromogenic (Panel B) Immunodetection kit (anti-mouse).
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