Ferric Nitrilotracetate - How to make it out of FeCl3 and Na NTA (Nov/14/2008 )
Sorry if the topic is in the wrong spot, but I wasn't sure where to put it. I really hope that one of you can help me out.
I need to make Ferric Nitrilotriacetate (NTA) out of FeCl3 and Na NTA and the only info I have (that's in a paper) is that these to are prepared to Fe NTA in a molar ratio 1:4. Nothing else and since I'm mostly clueless about anything chemical, I've come for your help.
With this info do you think I should just add the 2 in water (or PBS or other suggestions?) in a molar ratio from 1:4 and will the chemicals do the work for me when they're in solution.
It would be great to hear any thoughts on this.
Thanks in advance.
prepare the nta as the sodium form then add fecl3. the fe will exchange with the sodium.
is this in free solution or is the nta bound to a matrix?
if it is bound to a matrix, here is the recharging method (taken from the ni-nta superflow cartridge handbook, qiagen):
Regeneration by stripping and recharging
Stripping and recharging of Ni-NTA Superflow Cartridges is usually not
necessary. If an increase in back pressure or significant contamination of the
resin is observed, a cleaning-in-place procedure usually restores performance.
However, if performance is still not satisfactory, the Ni-NTA resin in the
cartridge can be stripped and recharged using the protocol below.
1. Strip the resin by washing with 10 column volumes of stripping
buffer (50 mM Na phosphate; 300 mM NaCl; 100 mM EDTA;
2. Wash the resin with 20 column volumes of deionized water.
3. Recharge the water-washed cartridge by loading 2 column volumes
of 100 mM NiSO4 (in deionized water). Salts of other metals
(chlorides or sulfates) may also be used.
4. Wash with 10 column volumes of deionized water and re-equilibrate
with 10 column volumes of Buffer NPI-10.
NPI-10* (Binding/lysis buffer for native conditions, 1 liter):
50 mM NaH2PO4 6.90 g NaH2PO4·H2O (MW 137.99 g/mol)
300 mM NaCl 17.54 g NaCl (MW 58.44 g/mol)
10 mM imidazole 0.68 g imidazole (MW 68.08 g/mol)
Adjust pH to 8.0 using NaOH and sterile filter (0.2 or 0.45 μm).
* 1% Igepal CA-630 (Nonidet P40) should be added to lysis buffer when preparing cleared
lysates from insect or mammalian cells.
The cartridge is now ready for use. Store cartridge in 20–30% ethanol or
10–100 mM NaOH.
you can substitute fecl3 for niso4.
Ok, late reply, but thanks.
I only need it in free solution. But thanks for the info, just in case I need it!