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Compound 21 (15 mg, ca. 10%%) appears to be unstable upon chromatography and cannot be isolated in pure form.
HpnC and HpnD were obtained as pure proteins upon chromatography on a Ni2+ column; however, HpnC lost a substantial amount of activity during chromatography.
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There was a close agreement between the results for HbA1c assays based on venous and DBS samples (DBS = 0.9858venous + 0.3809), except for assays based upon affinity chromatography.
In general, good enantioselectivities were achieved when R was an aromatic substituent, although the yields were low to moderate due to suspected instability of the products upon column chromatography.
Upon size-exclusion chromatography, PS2/AP complexes appear mainly to consist of dimeric reaction centers with a slightly larger Stokes radius than PS2/dodecylmaltoside complexes.
After purification, the fractions contained at least 98% pure protein in 10 mM Tris buffer at pH 7. Upon isoelectric focusing chromatography, the recombinant CA3427 protein showed a PI≈5.
This material behaved similarly to the CDG Ig EBV cell derived material upon QAE Sephadex chromatography, was neutralised with alkaline phosphatase to yield predominantly Man5GlcNAc2 (Fig. 3C, lane 2) but, as expected from the specificity of endoH, was insensitive to this enzyme (results not shown).
We detected a significantly earlier elution of pParkin upon size-exclusion chromatography, suggesting an increased hydrodynamic size (EV2D).
However, there is currently no evidence supporting formation of tetrameric EhRacC/EhPAK4 in solution; e.g., the migration of EhRacC/EhPAK4 upon gel filtration chromatography was consistent with a 1 1 rather than a 2 2 complex.
Upon ion exchange chromatography on SP-Sepharose, fraction C3 was resolved into a large unadsorbed fraction S1 and two smaller adsorbed fractions (S2 and S3) of approximately the same size.
The resulting mutant (Ficolin-B PO) was expressed and purified from CHO cells and resembled WT protein upon gel filtration chromatography and SDS-PAGE (data not shown), confirming that it assembles correctly during biosynthesis.
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