Western Blot of L1 worms

Model Organism: Worms Methods & Reagents
1

Hi

I am quite new to this forum so please forgive me if I violate any laid down rule (I’m assuming there are quite a few)

I intend to do a western blot (WB) on L1 worms in order to quantify SNA-2, SNA-1 and SUT-1 proteins in a strain that I made. The problem is that I do not know how many worms I will have to pick for this assay so as to get a good signal.

Has anyone done a WB on L1 worms before? How many worms will you suggest I pick, to get a good signal (assuming the primary antibody I want to use is already optimised)?

I will appreciate any information that will make my assay go smoothly, as this is the first time I will be doing this assay.

2

Hi,

The number of worms depends on the expression level of your protein and the sensitivity and specificity of the antibody. In my past experience, I need to pick 20-30 L1 worms per load. The best antibody only needs a single worm, especially for the adult.
However, I prefer to bleach the worms to get a full plate of the eggs. After they hatch, wash them off. Spin them down with low speed (about 1000rpm, 1min). Repeat the wash and spin to remove the bacteria.