Anti-6X His tag® antibody [AD1.1.10] (ab15149)

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Overview

  • Product name
    Anti-6X His tag® antibody [AD1.1.10]
    See all 6X His tag® primary antibodies
  • Description
    Mouse monoclonal [AD1.1.10] to 6X His tag®
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, IHC-Fr, IP, WBmore details
  • Species reactivity
    It reacts against all His tagged proteins so far tested. In Western blotting of bacterial extracts the antibody has been shown not to react with any endogenous products, although some Reactivity may be seen with extracts of insect or mammalian cells.
  • Immunogen

    PAX6 transcription factor linked to histidine tag.

  • General notes

    HIS-TAG® is a trademark of EMD Biosciences, Inc.

     

    ab15149 recognises proteins containing the motif HHHHHH and is therefore of value in detecting proteins containing histidine tags. The antibody may be used to track the purification of proteins expressed in bacteria, yeast or insect cells.

    The clone number has been updated from (3H2201) to (AD1.1.10) both clone numbers name the same antibody clone.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Protein G purified
  • Purification notes
    Tissue cuture supernatant purified by Protein G affinity chromatography.
  • Primary antibody notes
    ab15149 recognises proteins containing the motif HHHHHH and is therefore of value in detecting proteins containing histidine tags. The antibody may be used to track the purification of proteins expressed in bacteria, yeast or insect cells.
  • Clonality
    Monoclonal
  • Clone number
    AD1.1.10
  • Myeloma
    NS1
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab15149 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/500 - 1/100.

Target

  • Relevance
    The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
  • Cellular localization
    Depends upon the localization of the parent protein tagged with hexahistidine.
  • Alternative names
    • 6 His epitope tag antibody
    • Hexa His tag antibody
    • HHHHHH epitope tag antibody
    • HHHHHH tag antibody
    • His tag antibody
    • Polyhistidine Tag antibody
    see all

References

This product has been referenced in:
See all 5 Publications for this product

Publishing research using ab15149? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Question

I was looking for an anti-His-tag antibody. I
found that in the description in some of them (ab5000, ab18184) it is
indicated that the antibody recognizes recombinant protein expressed
in mammalian cells and in some other (ab15149, ab81663) it is
specified that the antibody recognizes protein expressed in bacteria.
If the antibody is raised against His-tag epitope can it recognize
indifferently recombinant protein expressed either in bacteria either
in mammalian or eukariotic cells? or not?
I had some problem with an anti-His-tag antibody of another company
since recognize the protein expressed in bacteria but not in mammalian
in WB and I would like to understand the reason before to look for a
product from a different company.

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Answer

I can confirm that all four products, the ab5000, ab15149, ab18184 and ab81663 detect proteins, which contain either an N or C-terminal HHHHHHGS (6x) epitope tag.

Important aspects for his tag recognition are, the localisation of the tag (N- or C-terminal) and the length of the his tag, that should be identical with the antibody chosen.

Furthermore it is important to confirm that the tagged protein is expressed in transfected or transformed cells. If the recombinant protein is expressed, it should not make a difference for the detection if either a bacteria or a mammalian system was used.

However, some his tag antibodies could cross react with other proteins in the used system. For ab81663, ab15149 there may be seen some cross reactivity with extracts of insect or mammalian cells

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Question

1.does ab15149 recognize n terminal histidine tags ? 2. If so does require just 6x His or 6xHis plus something else at the n terminal for the recognition ?

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Answer

This antibody recognises proteins containing the motif HHHHHH (5x His) and it is therefore of value in detecting proteins containing histidine tags (either at the N- or C-terminal). The antibody may be used to track the purification of proteins expressed in bacteria, yeast or insect cells.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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