General description
His-Tag is a series of at least 6 histidine residues used as
a tag motif at the N- or C-terminus of recombinant proteins. The relatively
small size of this tag vs. other commonly used tags such as GST (30 kDa) makes
it less likely to interfere with target protein structure or function. Its
small size also makes it more suitable for use under denaturing conditions. His
tags are most often used for affinity purification and binding assays. His tags
bind to nickel or cobalt ions with high affinity, and this interaction with
metal ions is the basis of many his-tag based affinity media. The His Tag
epitope is widely used to detect expression of recombinant proteins in
bacterial, yeast, insect,and mammalian systems.
Immunogen
6x His synthetic peptide.
Application
Immunocytochemistry:
2 μg/mL of a previous lot of antibody showed positive immunostaining for His
tagged Akt1/PKBα in transfected HeLa cells.
Immunoprecipitation:
This antibody has been reported by an independent laboratory to
immunoprecipitate His Tagged proteins.
Research Category
Epitope Tags & General Use
Research Sub Category
Epitope Tags
RNA Binding Protein (RBP)
This Anti-Histidine Tagged Antibody, clone HIS.H8 is
validated for use in IC, IP, WB for the detection of Histidine Tagged.
Biochem/physiol Actions
Recognizes tag sequence HHHHHH on either the N- or
C-terminus of any recombinant fusion protein; no other amino acids surrounding
the 6x-His tag sequence are required for the recognition of the 6x-His tag.
Physical form
Format: Purified
Purified
Purified mouse monoclonal IgG2b in buffer containing PBS, pH
7.4 with 0.05% sodium azide.
Preparation Note
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations:
Upon first thaw, and prior to removing the cap, centrifuge the vial and gently
mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid
repeated freeze/thaw cycles, which may damage IgG and affect product
performance.
Analysis Note
Control
RIPA lysates of transfected HeLa cells.
Routinely evaluated by western blot and immunocytochemistry
on transfected HeLa cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected His tagged Akt1/PKBα in RIPA lysates of
transfected HeLa cells.
Other Notes
Concentration: Please refer to the Certificate of Analysis
for the lot-specific concentration.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt,
Germany
Disclaimer
Unless otherwise stated in our catalog or other company
documentation accompanying the product(s), our products are intended for
research use only and are not to be used for any other purpose, which includes
but is not limited to, unauthorized commercial uses, in vitro diagnostic uses,
ex vivo or in vivo therapeutic uses or any type of consumption or application
to humans or animals.
