[ccp4bb] off topic: Is deglycosylation necessary for crystallization?

Hi Engin, Thanks for the correction. This saved a lot of fancying of mine for moving to insect cells :-). Let me also add a little on the yeast side (no personal empirical evidence either): wt yeasts generate hypermannosyl N-glycans, which are tens of to more than a hundred mannose (or m

[ccp4bb] immobilized DNA resin

Hi, I have never done this myself, but as far as I know, DNA can be directly conjugated through their primary amino groups to CNBr-activated beads or NHS-activated agarose beads. These beads are supplied by many companies: pierce, sigma, biorad, GE healthcare, etc.. - the same thing used for m

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

John, I believe my statement is accurate. The Compton & Jones paper you cite states in the abstract: "Interactions are chiefly with arginine rather than primary amino groups; the other basic (His, Lys) and aromatic residues (Try, Tyr, and Phe) give slight responses. The binding behavior is attr

Re: [ccp4bb] immobilized DNA resin

Hi Alex, Most DNA-binding proteins has decent affinity to heparin columns. Have you tried purifying your protein over heparin columns? Cheers, Raji On Sat, Apr 9, 2011 at 8:44 PM, Alexandra Deaconescu wrote: > Hello ccp4 enthusiasts: > > I am afraid this is a non-ccp4 related question. Can a

[ccp4bb] immobilized DNA resin

Hello ccp4 enthusiasts: I am afraid this is a non-ccp4 related question. Can anyone recommend an immobilized dsDNA chromatographic resin for purification of DNA-binding proteins? GE seems to have something - I was wondering if people have other recommendations? In the age of GST and His tags

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

It is not surprising that your bradford and BCA assays don't agree if you have no aromatic amino acids in your protein. Bradford dye binds to hydrophobic residues, mainly aromatics, so I would guess your bradford is consistantly giving lower measurements than the BCA assay. I also wouldn't be s

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

At 9:47 AM -0700 4/9/11, Michael Thompson wrote: Bradford dye binds to hydrophobic residues, mainly aromatics, The statement above is not accurate. Compton and Jones. Anal. Biochem. 151(2): 369-374, 1985 - John --

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

Michael Thompson a écrit : There is a very simple and very quick method that yields an answer approx. 15% reliable: measuring the increment of index of refraction due to the protein. The measurement of an index of refraction can be very accurate. You "only" need something like a 5µl drop

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

It is not surprising that your bradford and BCA assays don't agree if you have no aromatic amino acids in your protein. Bradford dye binds to hydrophobic residues, mainly aromatics, so I would guess your bradford is consistantly giving lower measurements than the BCA assay. I also wouldn't be su

Re: [ccp4bb] how to quantitate protein which dont have ne aromatic residue

you can do amino acid analysis on your pure protein, using a commercial or academic service - I hope these are still around. You should only need to do this once, then relate the result to your A220, BCA and Bradford assays. Mark Quoting Arpit Mishra: > hello everybody > > i am working on the

[ccp4bb] how to quantitate protein which dont have ne aromatic residue

hello everybody i am working on the protien which dont have any aromatic residue i do fplc other purification using 220 absorption, but i want to quantitate protein precisely i have tried using BCA nd bradford but both methods quantification is not matching,,so any one is having sum idea how to q