Key Difference – Nitrocellulose vs Nylon Membrane
Blotting is an important technique for the detection of specific sequences of DNA, RNA, and proteins from their mixtures in molecular biology. It is done using a membrane called blot. There are different blotting techniques such as northern, southern and western blotting. Selecting a proper membrane for the blotting process should be carefully done to prevent nonspecific binding and wrong detections. Nitrocellulose, nylon, and PVDF are commonly used membranes in blotting techniques. They have different characteristics. The key difference between nitrocellulose and nylon membrane is that nitrocellulose membranes have high protein immobilization potential whereas nylon membranes have high nucleic acid immobilization potential. However, both types of membranes are frequently used in blotting techniques.
CONTENTS
1. Overview and Key Difference
2. What is a Nitrocellulose Membrane
3. What is a Nylon Membrane
4. Side by Side Comparison -Nitrocellulose vs Nylon Membrane
5. Summary
What is a Nitrocellulose Membrane?
Nitrocellulose membrane is a commonly used membrane filter in nucleic acid and protein blotting techniques. It has a high protein binding potential. Hence, nitrocellulose membranes are widely used in western blotting technique. Nitrocellulose membranes are compatible with all hybridization techniques and show superior binding capacities without interferences. Nitrocellulose membranes are naturally hydrophilic in nature. They make hydrophilic interaction with molecules and efficiently immobilize them onto the membrane. Commercial nitrocellulose membranes are available in two pore sizes: 0.45 and 0.2 µm.
Figure 01: Nitrocellulose membrane used in western blotting
What is a Nylon Membrane?
Nylon membrane is another type of commercial membrane used in blotting techniques. It is alternatively used with nitrocellulose membranes for southern and northern blotting. Nylon membranes are ideal for southern blotting than nitrocellulose due to their high affinity to binding with DNA. Due to several unique features of nylon membranes, researchers commonly use nylon membranes for southern and northern blotting instead of over nitrocellulose membrane. Nylon membranes are also recommended for stripping and reprobing, unlike nitrocellulose.
Figure 02: Nylon membrane used for southern blotting
What is the difference between Nitrocellulose and Nylon Membrane?
Nitrocellulose vs Nylon Membrane |
|
| Nitrocellulose membranes are brittle. | Nylon membranes are less brittle. |
| Handling | |
| They are difficult to handle. | They are easy to handle. |
| Reprobing | |
| Unsupported nitrocellulose membranes are hard to reprobe. | Reprobing is easy with nylon membranes. |
| Compatibility with Various Storage Conditions | |
| Nitrocellulose membranes have less strength to withstand various storage conditions. | Nylon membranes respond more robustly to various storage conditions. |
| Use | |
| Prewetting is required. | Prewetting is not required for nylon membranes. |
| Hydrophilic Nature | |
| Nitrocellulose membranes are hydrophilic in nature but are less hydrophilic than nylon membranes. | They are highly hydrophilic in nature. |
| Immobilization Potential | |
| Nitrocellulose membranes have less affinity for nucleic acids. But it has a high affinity for proteins. | Nylon membranes have a high binding potential with nucleic acids than nitrocellulose membranes. |
Summary – Nitrocellulose vs Nylon Membrane
Nitrocellulose and nylon membranes are special sheets used in blotting technique to reproduce the banding pattern on the gel. They enable the possibility of detecting a specific sequence or a protein from the mixtures by immobilizing them on the membrane. Once the molecules immobilized on the membrane, it can be used as a substrate for hybridization analysis with labeled probes. Nitrocellulose membrane is commonly used for protein detection in western blotting technique due to its high binding affinity with proteins. Nylon membranes are often used for southern and northern blotting. This is the difference between nitrocellulose and nylon membrane.
References:
1. Hayes, P. C., C. R. Wolf, and J. D. Hayes. “Blotting techniques for the study of DNA, RNA, and proteins.” BMJ: British Medical Journal. U.S. National Library of Medicine, 14 Oct. 1989. Web. 07 Apr. 2017
2. Mahmood, Tahrin, and Ping-Chang Yang. “Western Blot: Technique, Theory, and Trouble Shooting.” North American Journal of Medical Sciences. Medknow Publications & Media Pvt Ltd, Sept. 2012. Web. 07 Apr. 2017
Image Courtesy:
1. “Western blot transfer” By Bensaccount at English Wikipedia (CC BY 3.0) via Commons Wikimedia