Analysis Note

ControlRIPA lysates of transfected HeLa cells.

Application

Research Sub CategoryEpitope TagsRNA Binding Protein (RBP)

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 CategoryEpitope Tags & General Use

This Anti-Histidine Tagged Antibody, clone HIS.H8 is validated for use in IC, IP, WB for the detection of Histidine Tagged.

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.

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.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified

Purified mouse monoclonal IgG2b in buffer containing PBS, pH 7.4 with 0.05% sodium azide.

Format: Purified

Quality

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.

Specificity

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.

Storage and Stability

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.