Difference Between Nitrocellulose and PVDF

While both nitrocellulose and PVDF membranes are used for Western blotting and amino acid analysis, nitrocellulose is ideal in detecting low molecular weight proteins while PVDF is more suitable for detecting higher molecular weight proteins.

1. What is the difference between PVDF and nitrocellulose membranes?
2. What are the advantages and disadvantages of PVDF and nitrocellulose?
3. Why is nitrocellulose used in Western blot?
4. What is a PVDF membrane?
5. What does PVDF stand for?
6. Why do you soak PVDF membrane in methanol?
7. How do you keep PVDF membranes?
8. Can protein pass through PVDF membrane?
9. How do you activate the nitrocellulose membrane?
10. Why is glycine used in running buffer?
11. What is the pore size of nitrocellulose?
12. What is the principle of Western blot?

What is the difference between PVDF and nitrocellulose 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 are the advantages and disadvantages of PVDF and nitrocellulose?

3. What are the advantages and disadvantages of PVDF and nitrocellulose? PVDF is a highly nonreactive plastic that immobilizes proteins based on its non-specific affinity for amino acids. PVDF is preferred over nitrocellulose because it offers better protein retention, strength, and chemical compatibility.

Why is nitrocellulose used in Western blot?

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.

What is a PVDF membrane?

Polyvinylidene fluoride (PVDF) membranes bind biomolecules through hydrophobic interactions. Membrane pores increase the surface binding area while restricting sizes of bound proteins. ... It has a higher protein binding capacity and a higher retention than 0.45 µm membranes.

What does PVDF stand for?

PVDF is the abbreviation for polyvinylidene fluoride, which can also be referred to as PVF2. This polymer is part of a class of materials known as fluorocarbons or fluoropolymers, which are characterized by high thermal stability and excellent chemical resistance.

Why do you soak PVDF membrane in 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.

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.

Can protein pass through PVDF membrane?

Initial transfer performed overnight (16hr) at 30V failed because small and mid sized proteins (smaller than 100kDa) transferred right through the membrane (as evident from Ponceau staining of the blot and Coomassie staining of the gel).

How do you activate the nitrocellulose membrane?

They mentioned that : 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.

Why is glycine used in running buffer?

Glycine is in the running buffer, which is typically at a pH of 8.3. ... When an electric field is applied, glycinate anions hit the pH 6.8 stacking buffer, and change to become mostly neutrally charged glycine zwitterions. That means they move slowly through the stacking layer toward the anode due to their lack of charge.

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.

What is the principle of Western blot?

Western blotting (protein blotting or immunoblotting) is a rapid and sensitive assay for detection and characterization of proteins. It is based on the principle of immunochromatography where proteins are separated into polyacrylamide gel according to their molecular weight.