[Western Blotting] Diffusion of Antibodies in PVDF and NZ membranes

[Kyle Legate]

lowenbrau wrote:
>> newsnet customer wrote:
>>>> newsnet customer <customer from newsnet.com> wrote:
>>>>
>>>>> I have experienced protein diffusion from my membrane after an 
>>>>> overnight
>>>>> antibody incubation. This was confirmed with a positive control. And 
>>>>> since
>>>>> he had no signal or reduced signal on his membrane, this could be a 
>>>>> possible
>>>>> explanation. Please realise that comments on a newsgroup should be 
>>>>> taken
>>>>> with a grain of salt. And you should take a chill pill.
>>>> So, as Kyle suggested, you are a troll.  If your technique in the lab is
>>>> as thorough as your knowledge, there are better explanations for your
>>>> failed Western.
>>> Can you read troll? As confirmed my western failed because of the long 
>>> incubation time. However, i'm curious to hear some of your "better 
>>> explanations" especially when you don't know the details of my experiment 
>>> or the experiment of the original post. Tool.
>>>
>> I'm willing to bet you tried to blot your protein to nylon.
> 
> Many people do so whats ur point? and don't tell me that PVDF doesn't have 
> this problem cos ur a tool.
> 
Wow, a blast from the past. Actually, nucleic acids, not proteins, are 
blotted to nylon. And PVDF doesn't have the problems outlined above. 
Protein binding is essentially irreversible under conditions you would 
use in the lab. But thanks for the effort of replying, "ST".