MagneHis™ Protein Purification System Technical Manual
Literature # TM060
The MagneHis™ Protein Purification System provides a simple, rapid and reliable method for the purification of polyhistidine- or HQ-tagged proteins. Paramagnetic precharged nickel particles (MagneHis™ Ni-Particles) are used to isolate polyhistidine- or HQ-tagged protein directly from a crude cell lysate of bacterial, insect or mammalian cells or from culture medium using either a manual or automated procedure. Using the manual protocol, polyhistidine- or HQ-tagged protein can be purified using 1ml of culture of up to 6 O.D.600 of bacterial cells, or 2 × 106 insect or mammalian cells. Samples can also be processed using a robotic platform for high-throughput applications. The MagneHis™ System is useful for screening multiple clones for expression, optimizing expression conditions (temperature, media, host strain, etc.) and the initial screening of mutant clones.
Bacterial, insect or mammalian cells containing a polyhistidine- or HQ-tagged protein can be lysed directly in the culture medium using the provided FastBreak™ Cell Lysis Reagent, 10X, or by adding 1X FastBreak™ Cell Lysis Reagent to cell pellets. MagneHis™ Ni-Particles are then added to the lysate. In addition, tagged proteins secreted into the medium of insect or mammalian cells can be purified without interference from serum in the medium. Polyhistidine- or HQ-tagged proteins bind to the MagneHis™ Ni-Particles in a matter of minutes. Unbound proteins are washed away, and the target protein is recovered by elution with imidazole. Polyhistidine- or HQ-tagged proteins can be purified under native and denaturing conditions. Additionally, HQ-tagged proteins may elute with a lower concentration of imidazole (50–100mM) compared to polyhistidine-tagged proteins.
Printed in USA. Revised 8/09.