Overview

  • Product name
    Anti-6X His tag® antibody - ChIP Grade
    See all 6X His tag® primary antibodies
  • Description
    Rabbit polyclonal to 6X His tag® - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    ELISA: Antibody specificity was verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA). A 1:25,000 dilution of the antibody gave an OD=1.0 in a 15 minute reaction using HRP-conjugated Goat anti-Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run.
  • Tested applications
    Suitable for: IP, WB, IHC-P, ICC/IF, ELISA, ChIP, Flow Cytmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Synthetic peptide: HHHHHH conjugated to KLH.

  • Positive control
  • General notes
    This antibody recognizes both C-terminal and N-terminal tags. HIS-TAG® is a trademark of EMD Biosciences, Inc.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

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
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9108 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
IP Use at an assay dependent concentration.
WB 1/1000.
IHC-P Use at an assay dependent concentration. PubMed: 21483675
ICC/IF 1/100 - 1/500.
ELISA 1/1000.
ChIP Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads.
Flow Cyt Use at an assay dependent concentration.

(see Abreview published on 22nd January, 2007).

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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

Images

  • ab9108 staining CACNB3 with His tag expressed in HEK293 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum, 0.1% TX100 and PBS) for 16 hours at 4°C. An  Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/750).

    See Abreview

  • A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).
  • All lanes : Anti-6X His tag® antibody - ChIP Grade (ab9108) at 1/4000 dilution

    Lane 1 : HeLa whole cell lysate - transfected with empty vector
    Lane 2 : HeLa whole cell lysate - transfected with His-tagged gene

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 35 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds

    See Abreview

  • Use of ab9108 in western blotting at 1:1000 against Epitope Tag Positive Control lysate (ab4524).
    Lane 1: 2 µl lysate
    Lane 2: 1 µl lysate
    Lane 3: 0.5 µl lysate
    Lane 4: 0.25 µl lysate Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate Lane 3: 0.5 µl lysate Lane 4: 0.25 µl lysate

References

This product has been referenced in:
  • Diakonov EE  et al. Evidences against vesicle-dependent trafficking and involvement of extracellular proteasomes into cell-to-cell communications. Biochem Biophys Res Commun 508:368-373 (2019). Read more (PubMed: 30503341) »
  • Müskens FM  et al. Design, Synthesis, and Evaluation of a Diazirine Photoaffinity Probe for Ligand-Based Receptor Capture Targeting G Protein-Coupled Receptors. Mol Pharmacol 95:196-209 (2019). Read more (PubMed: 30514721) »
See all 73 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

Unfortunately we do not have a His tagged BSA in our catalog and can not offer HIS-tagged BSA for sale as it's just a component within our QC procedures.

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Answer

Thank you for your enquiry. Ab9108 recognizes both C-terminal and N-terminal tags.

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Answer

Yes, this antibody recognizes both C-terminal and N-terminal tags.

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Answer

Sorry but we do not have this product in our catalogue. If you cannot find what you are looking for within the Abcam catalog, I suggest that you click on the link to The world's Antibody Gateway. The world's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogs of all online antibody companies (currently 249).

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