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His-probe (H-15): sc-803
Santa Cruz Biotechnology, Inc.
1.800.457.3801 831.457.3800 fax 831.457.3801 Europe
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Plasmid vectors for the expression of coding regions of eukaryotic genes in
bacterial, insect and mammalian hosts are in common usage; such expres-
sion vectors are frequently used to encode hybrid fusion proteins consisting
of a eukaryotic target protein and a specialized region designed to aid in the
purification and visualization of the target protein. A system that has proven
to be very successful relies on the insertion of a six histidine (His6)
sequence in the N-terminus of the encoded protein, allowing for efficient
coupling to Ni++-chelating resins and purification by single step affinity chro-
matography. This polyhistidine sequence can then be removed by specific
cleavage at sites recognized by enzymes such as thrombin or enterokinase,
permitting the separation of the target protein from the polyhistidine tag.
Visualization of such fusion proteins can be achieved by utilizing antibodies
generated against specific peptide sequences downstream from the multiple
1. Maniattis, T., et al. 1982. Molecular Cloning. Cold Spring Laboratory, Cold
Spring Harbor, NY.
2. Smith, D.B., et al. 1988. Single-step purification of polypeptides expressed
in Escherichia coli as fusions with glutathione S-transferase. Gene 67:
3. Hochuli, E. 1988. Large-scale chromatography of recombinant proteins.
J. Chromatog. 444: 293-302.
4. Thanos, D., et al. 1992. The high mobility group protein HMG I(Y) is
required for NF-κB-dependent virus induction of the human IFN-β gene.
Cell 71: 777-789.