His-Tag-Monoclonal Antibodies

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His-Tag

Qty


Total
$135
Catalog #
AE003
Antibody Type
Monoclonal Antibody
Gene ID
Swiss Prot
Size
Species
Mouse
Isotype
IgG
Purity
Affinity purification
Additional Information
Tested applicationsWB IF IP
Recommended DilutionWB 1:2000 - 1:5000 IF 1:50 - 1:500 IP 1:50 - 1:100
Observed MWRefer to Figures
ImmunogenA synthetic peptide of His tag
Storage BufferStore at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.
Images
  • AE003: image 1

    Western blot analysis of over-expressed His-tagged protein in 293T cell using His antibody at different dilution. Each lane was loaded with 2 ug cell lysate.

  • AE003: image 2

    Immunoprecipitation of over-expressed His-tagged protein in HeLa cell incubated using His antibody. A mock served as negative control and over-expressed HeLa cell lysate served as positive control.

Background

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.

Protocol

N/A

MSDS
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