1 reply to this topic

[Epigeneticist]

Hi,

I recently did a western with a new antibody. After developing the film, the protein marker showed up and it was a very strong signal. However, the samples only produced faint bands. I am currently probing for actin after stripping the membrane. I have never seen the marker produce a signal on a western before, any answers for the cause of this?

Thanks

[GNANA]

just reduce the quantity of marker you loading, you can reduce its signal, alternatively if you have extra well just skip a well and start loading the samples...may be your target antibody might not be picking signal in a short time, so your longer incubation and more marker loaded are the possible reason for this.. definitely your actin shd be good, so you wnt get signal dominance frm the marker...