Ni-chelate chrom.

Product: Rat recombinant calbindin D-9k (recombinant) produced in Escherichia Coli. Lot No.: 5 Reconstitution 10 µg protein reconstituted with 25 µl PBS (0.4 µg/µl) 60 µg protein reconstituted with 150 µl PBS Storage Freeze in small aliquots (e.g. 1 or 10 µg) and keep at -80°C. Avoid repeated freezing and thawing. Purification E. coli containing a His6-fusion protein were disrupted in a buffer containing 6M GuHCl/0.1 M NaH2PO4. After a low speed centrifugation to remove cell debris, the supernatant was applied to the charged NTA resin column, equilibrated in disruption buffer, before the column being washed with the same buffer. Afterwards, the column was washed and the HIS6- fusion proteins eluted by decreasing the pH of the eluent (8 M Urea /0.1 M Na2HPO4) in a stepwise manner (pH 6.3 / 5.9 / 4 / 2). The protein was dialysed against phosphate buffered saline, 0.1 M, pH 7.3 (PBS).