cableties的部落格

1,How to identify NC membrane filter, PVDF membrane filter, nylon membrane filter?

Nylon membrane is the ideal nucleic acid solid support, there are many types; Nitrocellulose (CN) membrane is currently the most widely used solid support, the cheapest price; PVDF membrane between the Nylon and CN membrane filter .

2,Membrane Protein Binding capacity

• In combination with DNA and RNA, RNA binding capacity of up to 480-600μg / cm2, can be combined with as short as 10bp nucleic acid fragments;
• Nitrocellulose membrane binding DNA and RNA capacity of up to 80-100μg / cm2, for 200bp DNA binding capacity is not strong;
• PVDF membrane binding DNA and RNA capacity of up to 125-300μg / cm2.

3,In the case of temperature suitability

the partial pyrimidine base in the nucleic acid can be bound to the positive charge on the membrane after baking or ultraviolet irradiation;
The cellulose membrane relies on hydrophobic interaction to bind DNA, which is not strong; PVDF Membrane binding is firm, resistant to high temperatures, and is particularly suitable for western blotting.

4,in terms of toughness

• Nylon membrane filter is strong;
• Nitrocellulose membrane filter is more brittle, easy to break;
• PVDF membrane filter stronger.

5,In the case of repeatability

• Nylon membranes can be repeatedly used for molecular hybridization, and the probe molecules can be eluted by alkalinity after hybridization;
• NC membranes can not be reused;
• PVDF membranes can be reused.

Q:When doing Western Blot, PVDF membrane with methanol soaking the purpose?
A:The purpose of the PVDF membrane with methanol foam is to activate the positive group above the PVDF membrane, making it easier to combine with negatively charged proteins and the addition of methanol for small molecule protein transfer is also the goal.