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Western Blotting substrate - Lumi-Light Western Blotting Substrate (Apr/19/2006 )

Has anyone had any experience using Lumi-Light Western Blotting Substrate (Roche)? I used to use Lumi-Light PLUS Western Blotting Substrate, but now we switched. I still use Protran BA 85 Nitrocellulose (Whatman) membranes and my blots used to be just fine. But now, the signal is down to no signal at all. And I just don't understand why. As far as I can tell, I don't do anything differently compared to before. Except of course, the substrate. But I can't imagine that that might make such a big difference... has anyone had similar experiences?

-das_schnaf-

what does Roche say about the difference between the two?

perhaps the 'plus' is just much more sensitive? have you tried using a little more?

-aimikins-

QUOTE (aimikins @ Apr 19 2006, 07:13 PM)
what does Roche say about the difference between the two?

perhaps the 'plus' is just much more sensitive? have you tried using a little more?



Yes, it is less sensitive. 1 pg of antigen is sufficient to be detected by the Plus compared to 10-50 pg by the normal one. I was just wondering if that makes such a big difference. The problem is I don't know how much antigen I have since I am using cell lysates. I am doing another blot now in order to compare the two substrates. I do hope that will solve it.

-das_schnaf-

Ok. Problem solved. The developer wasm't working properly. blink.gif

-das_schnaf-

Hi,
dont worry too much, in research this saying is quite common.

BARKING UP THE WRONG TREE

try to avoid the situation by pointing out the most probable cause, that is all we can do.

gud luk

QUOTE (das_schnaf @ Apr 20 2006, 07:35 AM)
Ok. Problem solved. The developer wasm't working properly. blink.gif

-payeli-