Words, Words, Words

Words, Words, Words - it's only words (Jul/21/2009 )

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fuse

but with experience, you'd get better at predicting and then you do what you have to do....are we still talking about the canadian rockies? ... :lol:

-casandra-

fuselage

who knows... a seismometer is a good idea...and noticing the smell of sulphur...

-hobglobin-

debris

now you're learning... :lol:
and I've to go now- to brave the snow and ice....and you have a good night's rest my darling clueless as always old dude...sleep tight and don't let the volcanic eruptions wake you....

-casandra-

space

then have a safe trip home on your adventurous trip over several volcano calderas. Good night darling the human volcano.

-hobglobin-

the final frontier

vetticus3 on Tue Feb 8 10:19:36 2011 said:

ex

we are a virus, and the earth is our host.

Poetic.

-lab rat-

médecins sans frontières

hey guys...hi LaRa and V is inspired by her fishies....

I'm lighting some candles now, Ameya (this name has a meaning?)....

-casandra-

doctors without borders

no... that was highschool philosopy.

-vetticus3-

scrubs

hi guys, apparently protein extraction was succesful!

hey v, technical question out of the biochemistry subforum... do you use sds in your extraction buffer? if so, what's the amount you use? i was using 1% but it seemed to interfere when i tried to quantify it with bradford reagent, then decreased it to 0.5% and i hope that won't affect the extraction quality? :unsure:

-toejam-

Dr. Percival "Perry" Ulysses Cox

sds interferes with bradford. for westerns, i add 1%, and don't measure the protein level. i usually count the cells, and use a control band (ie beta actin). we also had a nanodrop that could measure protein in the presence of sds by the end. there is also a kit from pierce that can measure protein in up to 5% sds (?).
i'll post this, and see what i can find about that kit.
there are other extraction buffers too... if you used ripa, i think that has 0.1% sds. there is also a kit that can extract protein using a column that has no sds (i think this is pierce). millipore also has a kit to remove detergent out of protein solutions, it's called something like detergent out.

- EDIT

quite a few companies have alternative to bradford that are compatible with sds. invitrogen has a bunch of them - they're usually good about giving out samples too.
ripa has 0.1% sds, so maybe that could be an option for you.
millipore's product is "detergen-OUT" tm. "The Chemicon Detergent-OUT SDS Kit is a simple high performance method for removing detergents from protein solutions. Removes detergents without significant dilution of protein solution. Simply load the protein solution on the Detergent-OUT Spin column and collect detergent free protein solution. Micro columns are suitable for removing detergent from a maximum of 0.2 mL sample volume."

-vetticus3-

Dr Bob Kelso

wow, thanks v! 5% sds sounds crazy! in case the problem persists i'll check ripa out

-toejam-