His60 Ni Magnetic Beads
His60 Ni Magnetic Beads
Brand: Takara Bio.
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His60 Ni Magnetic Beads
High-throughput his-tagged microscale purification—His60 Ni Magnetic Beads
His60 Ni Magnetic Beads provide the selective chemistry of His60 Ni immobilized metal affinity chromatography (IMAC) resin in a magnetic bead format. The beads are designed for the microscale purification of high and low molecular weight recombinant his-tagged proteins under native or denaturing conditions, when placed on a magnetic separator.
Overview
- Quick and easy separation of his-tagged proteins
- Selective His60 Ni chemistry for increased purity
- Eluted samples ideal for small volume applications
Applications
His60 Ni Magnetic Beads provide simple, effective separation of recombinant his-tagged proteins for a wide range of applications, including:
- Microscale purification of his-tagged proteins
- Studying protein structure and function
- Preparing proteins for X-ray crystallography
- Performing assays that detect protein-protein and protein-DNA interactions
- Immunization to raise antibodies against a protein of interest
- Screening for protein expression
- Optimizing purification conditions for scale-up with His60 Ni Resin
His60 Ni Magnetic Beads can also be used to immobilize 6xhis-tagged proteins for affinity chromatography, in order to:
- Analyze protein-protein and protein-nucleic acid interactions
- Purify untagged subunits or nucleic acids that interact with the immobilized protein
- Perform antibody purification
- Study interactions between ligands and receptors
Advantages
- Choice of native or denaturing conditions to purify proteins
- Directed presentation of 6xHis-tagged proteins increases reproducibility and signal-to-noise ratios
- Varying the amount of beads per well allows for a wide range of binding capacities
- A powerful tool for analyzing interactions between biomolecules
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under native conditions
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under native conditions. 400 µl of clarified lysate from E. coli cells expressing AcGFP1-6xHis was added to His60 Ni Magnetic Beads and allowed to bind for 30 minutes. The beads were washed 3X with 500 µl of Wash Buffer, and eluted 4X with 100 µl of Elution Buffer (from the His60 Ni Buffer Set, Cat. No. 635655). A 10 µl aliquot of each sample was analyzed using a 12% SDS-PAGE gel. Lane M: Spectra Multicolor Broad Range Protein Ladder. Lane 1. Original sample. Lane 2. Flowthrough. Lane 3. Wash 1. Lane 4. Wash 2. Lane 5. Wash 3. Lane 6. Eluate 1. Lane 7. Eluate 2. Lane 8. Eluate 3. Lane 9. Eluate 4.
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under denaturing conditions
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under denaturing conditions. 500 µl of clarified lysate from E. coli cells expressing AcGFP1-6xHis was added to His60 Ni Magnetic Beads and allowed to bind for 30 minutes. The beads were washed 3X with 500 µl of Equilibration Buffer and 1X with 500 µl of Wash Buffer—and eluted 3X with 50 µl pf Elution Buffer (from the His60 Ni Buffer Set, Cat. No. 635655). 8M urea was added to all the buffers. A 10 µl aliquot of each sample was analyzed using a 4–20% SDS-PAGE gel. Lane M: Spectra Multicolor Broad Range Protein Ladder. Lane 1. Original sample. Lane 2. Flowthrough. Lane 3. Wash 1. Lane 4. Wash 2. Lane 5. Wash 3. Lane 6. Wash 4. Lane 7. Eluate 1. Lane 8. Eluate 2. Lane 9. Eluate 3.
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Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under native conditions
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under native conditions. 400 µl of clarified lysate from E. coli cells expressing AcGFP1-6xHis was added to His60 Ni Magnetic Beads and allowed to bind for 30 minutes. The beads were washed 3X with 500 µl of Wash Buffer, and eluted 4X with 100 µl of Elution Buffer (from the His60 Ni Buffer Set, Cat. No. 635655). A 10 µl aliquot of each sample was analyzed using a 12% SDS-PAGE gel. Lane M: Spectra Multicolor Broad Range Protein Ladder. Lane 1. Original sample. Lane 2. Flowthrough. Lane 3. Wash 1. Lane 4. Wash 2. Lane 5. Wash 3. Lane 6. Eluate 1. Lane 7. Eluate 2. Lane 8. Eluate 3. Lane 9. Eluate 4.
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under denaturing conditions
Purifying his-tagged AcGFP1 from a cell lysate using His60 Ni Magnetic Beads under denaturing conditions. 500 µl of clarified lysate from E. coli cells expressing AcGFP1-6xHis was added to His60 Ni Magnetic Beads and allowed to bind for 30 minutes. The beads were washed 3X with 500 µl of Equilibration Buffer and 1X with 500 µl of Wash Buffer—and eluted 3X with 50 µl pf Elution Buffer (from the His60 Ni Buffer Set, Cat. No. 635655). 8M urea was added to all the buffers. A 10 µl aliquot of each sample was analyzed using a 4–20% SDS-PAGE gel. Lane M: Spectra Multicolor Broad Range Protein Ladder. Lane 1. Original sample. Lane 2. Flowthrough. Lane 3. Wash 1. Lane 4. Wash 2. Lane 5. Wash 3. Lane 6. Wash 4. Lane 7. Eluate 1. Lane 8. Eluate 2. Lane 9. Eluate 3.
