Difference Between Nitrocellulose and Nylon Membrane

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.

1. What is the difference between nitrocellulose and PVDF membranes?
2. What is nitrocellulose membrane?
3. Why nitrocellulose membrane is used in Southern blotting?
4. How do proteins bind to nitrocellulose membranes?
5. Why is PVDF membrane activated by methanol?
6. What is the pore size of nitrocellulose?
7. How do you use nitrocellulose membrane?
8. What is nitrocellulose used for?
9. How do you keep PVDF membranes?
10. Why is DNA transferred to a nylon membrane?
11. Is Southern blot still used?
12. What are the steps in Southern blotting?

What is the difference between nitrocellulose and PVDF membranes?

PVDF membranes have a higher protein binding capacity than nitrocellulose. The protein binding capacity of PVDF ranges from 150-200 µg of protein/cm² and nitrocellulose ranges from 80-100 µg of protein/cm². Although PVDF has a higher binding capacity, it could result in increased background in some circumstances.

What is nitrocellulose membrane?

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.

Why nitrocellulose membrane is used in Southern blotting?

In the original protocol nitrocellulose membrane have been used for the blotting in case of Southern blot but in recent times nylon membranes have been implemented for the blotting process due to their ability to bind more amount of DNA efficiently which allows the Southern blot to be carried out with less amount of ...

How do proteins bind to nitrocellulose membranes?

Protein molecules bind to nitrocellulose membranes through hydrophobic interactions while molecules bind to PVDF membranes through hydrophobic and dipole interactions.

Why is PVDF membrane activated by methanol?

A short rinse (15-30 seconds) in methanol (or other 100% alcohol (ethanol or isopropanol)) prior to Western transfer will "hydrate" the membrane and allow improved transfer and protein binding. Nitrocellulose membranes are hydrophilic so can be fully hydrated by aqueous buffers.

What is the pore size of nitrocellulose?

Bio-Rad's 0.2 µm pore–size nitrocellulose membrane is dense 100% nitrocellulose. Pure nitrocellulose is the most frequently used membrane for the transfer of both proteins and nucleic acids.

How do you use nitrocellulose membrane?

Nitrocellulose—Place the membrane directly into a shallow dish containing 50 ml of 1X Transfer Buffer for several minutes. Filter paper—Soak the filter paper briefly in 1X Transfer Buffer immediately prior to use. Gel—Use the gel immediately following the run.

What is nitrocellulose used for?

Guncotton, or nitrocellulose (also known as trinitrocellulose and cellulose nitrate) is a mild explosive, used in rockets, propellants, printing ink bases, leather finishing, and celluloid (a mixture of nitrocellulose and camphor; first used to manufacture billiard balls).

How do you keep PVDF membranes?

To store protein-transfered NC or PVDF membrane for a longer time, it is better to dry the membrane and then keep at -20^oC without folding. For PVDF membrane, just rinse in methanol before you blot.

Why is DNA transferred to a nylon membrane?

Southern Blots

1). The blotted DNA is usually covalently attached to the nylon membrane by briefly exposing the blot to UV light. Transferring the DNA to the sturdy membrane is necessary because the fragile gel would fall apart during the next two steps in the process.

Is Southern blot still used?

Today Southern blotting has been largely replaced by real-time PCR to answer the same experimental questions. Real-time PCR detects and quantifies a gene or DNA sequence of interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard PCR.

What are the steps in Southern blotting?

Step-by-Step Guide to Southern Blot Analysis

1. Step 1 DNA digestion.
2. Step 2 Gel electrophoresis.
3. Step 3 Blotting.
4. Step 4 Probe labeling.
5. Step 5 Hybridization & washing.
6. Step 6 Detection.