Sure, we can talk about this off line.
-- Kevin
On Thu, Mar 8, 2012 at 7:48 PM, Pius Padayatti wrote:
> Kevin,
> thanks.Sine this is of not much interest to many here
> please feel free to go off line in the future.
>
> The method of dialysis is confusing here.
> How can one achieve this by b
Kevin,
thanks.Sine this is of not much interest to many here
please feel free to go off line in the future.
The method of dialysis is confusing here.
How can one achieve this by buffer exchange?
I wonder if an extensive wash of protein on a column would work
instead.
Pius
On Thu, Mar 8, 2012 at
In my case, this method was developed for large quantity sample
(~100g/batch) purification.
Under cGMP, I could not introduce EDTA or other chemicals like
Triton-X100 into the system. Otherwise, I just solve small problem but
bring into an even large problem to the manufacture line.
I hope you ca
To Pius,
In my case, the protein/biopolymer is Lysine/amine riched.
1. Lys or Arg are mainly positively charged under pH 7.0 ~ 8.0, and
then provide positively charge to LPA. Of course, some metal cations
could also be involved in LPA binding.
2. Tris with the NH2 group could also be protonated
This is in response to a comment to this thread
Kevin,
Could you explain how that worked?
How do you know your method worked?
Did you estimate the lipopolysaccharide before and after the method?
The method already mentioned here to wash using TritonX100 makes sense.
by washing bound protein sample
Dear Jerry,
I've just become aware of this spin column to remove endotoxin, marketed by
Generon, UK.
http://generon.co.uk/index.php?route=product/category&path=238
Good luck,
Jon
On Wed, Mar 7, 2012 at 9:50 AM, Jerry McCully
wrote:
>
>
> Dear All;
>
>
I tried to use 250mM Tris pH 7.6, 1M NaCl and 5~10% 2-propanol for
dialysis. It works,
On Wed, Mar 7, 2012 at 6:50 AM, Jerry McCully
wrote:
>
>
> Dear All;
>
> We purified a His tag protein by Ni-NTA and gel-filtration from
> E.coli.
>
> We tried two endotoxin removal resins fr
Hi Jerry,
We have used 0.05% Triton X-100 in our wash buffer to wash the protein
on column (Ni-IMAC for His-tagged proteins and MabSelect for
antibodies). 20 column-volumes wash followed by 20 column-volume wash
without detergent is typically enough to reduce the endotoxin
significantly.
I would al
Dear All;
We purified a His tag protein by Ni-NTA and gel-filtration from E.coli.
We tried two endotoxin removal resins from Pierce. However, it is hard
to remove the endotoxins in the purified protein because the protein bound to
the resin as well.
This p
