Ni-NTA Magnetic Agarose Beads (6 x 1 ml)

Protein tag Purification of His-tagged proteins

Experiment
Protein tag Purification of His-tagged proteins
Product
Ni-NTA Magnetic Agarose Beads (6 x 1 ml) from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
The manufacturer-estimated binding capacity of 20 µL of bead suspension (40 µg) was estimated to be in great excess of the amount of HRPII present

Publication protocol

Samples were prepared by spiking whole blood with D6 P. falciparum parasite culture to a final concentration of 200 parasites/µL, in order to closely mimic a patient sample. The blood was lysed at a 1:1 ratio with lysis buffer, and 50, 100, 200, or 500 µL volumes of lysed blood plus 20 µL of Ni–NTA magnetic particles were added to 1.5 mL Eppendorf tubes (n = 3 for each time point). The manufacturer-estimated binding capacity of 20 µL of bead suspension (40 µg) was estimated to be in great excess of the amount of HRPII present. The tubes were placed on a bench top vortexer fitted with a tube holder attachment and vortexed for 5, 10, 15, 30, or 60 min at medium speed. For each time point, three tubes from each sample volume were spun down on a minicentrifuge for 10 s to collect the sample and beads at the bottom of the tube. The tubes were then placed on a magnetic tube rack to pull the magnetic particles to the side of the tube while the blood sample supernatant was collected. The blood supernatant samples for each sample volume at each time point were analyzed by ELISA for HRPII content, using a previously published method [19].

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Manufacturer protocol

Download the product protocol from Qiagen for Ni-NTA Magnetic Agarose Beads (6 x 1 ml) below.

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