Plasmid vectors for the expression of coding regions of eukaryotic genes in
bacterial, insect and mammalian hosts are in common usage; such expression
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 chromatography. 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 cloning site.

1. Maniattis, T., et al. 1982. Molecular Cloning. Cold Spring Laboratory, Cold Spring Harbor, NY.
2. Hochuli, E. 1988. Large-scale chromatography of recombinant proteins. J.Chromatog. 444: 293-302.

This Abmart monoclonal antibody is produced by immunizing mice with a 6×His synthetic peptide (KLH-coupled).

His-Tag (10E2) Mouse mAb detects over-expressed or recombinant proteins
containing the 6× His epitope tag.

Store at -20°C. Stable for one year from the date of shipment.

All
 

Mouse IgG1

Western blotting                 1:5000
Immunofluorescence        1:1000
Immunoprecipitation          1:100
ELISA                                     1:2000