Overview

  • Product name
    Anti-HA tag antibody - ChIP Grade
    See all HA tag primary antibodies
  • Description
    Rabbit polyclonal to HA tag - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    ELISA: The anti HA diluted 1:70.000 gave an O.D.=1.0 in a 15 minute reaction against peptide conjugated with a different carrier than used for anti peptide purification. HRP conjugated Goat anti rabbit IgG was used and TMB was the substrate.
  • Tested applications
    Suitable for: ChIP/Chip, IP, ELISA, WB, ICC/IF, ICC, Flow Cyt, ChIPmore details
  • Immunogen

    YPYDVPDYA (influenza hemagglutinin-HA-epitope) conjugated to KLH.

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
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antibodies were immunoaffinity purified using the peptide conjugated to a solid-phase support.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab9110 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
ChIP/Chip Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ELISA 1/200 - 1/500.
WB 1/4000 - 1/10000.
ICC/IF Use a concentration of 1 - 4 µg/ml.
ICC Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

ChIP Use 3 µg for 25 µg of chromatin.

Target

  • Relevance
    Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag is derived from the HA molecule corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the proteins.
  • Alternative names
    • HA epitope tag antibody
    • HA1 antibody
    • HA2 antibody
    • hemagglutinin antibody
    • Hemagglutinin HA1 chain antibody
    • Hemagglutinin HA2 chain antibody
    see all

Images

  • All lanes : Anti-HA tag antibody - ChIP Grade (ab9110) at 1/4000 dilution

    Lane 1 : 15ug untransfected wcl lysate
    Lane 2 : 293FT cells transfected with 15kDa HA tagged Vpr (an HIV1 accessory protein)

    Secondary
    All lanes : HRP conjugated Goat anti-Rabbit

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 seconds


    This image is courtesy of an Abreview

    See Abreview

  • ab9110 was diluted to 4 µg/mg lysate and incubated with a nuclear lysate of HEK293T cells transiently expressing HA-tagged protein and a Protein A matrix for 2 hours a 23°C to achieve immunoprecipitation.  1000 µg of lysate was present in the input. 

    A HRP-conjugated anti-rabbit HA monoclonal antibody diluted 1/1000 was used for the Western Blot step.

    See Abreview

  • This image was kindly supplied as part of the review submitted by Kasper Fugger. Immunofluorescence staining of U-2 cells expressing HA-tagged protein with ab9110.
  • Xenopus laevis oocytes were injected with mRNA for HA-tagged human BORIS. Chromatin was prepared according to the Abcam X-ChIP protocol. Oocytes were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 20µl of Protein A/G sepharose beads, and 3µg of ab9110 (anti-HA, light blue) or, 3µg of ab18337 (anti-Boris, dark blue). A non-specific antibody was used as a control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

  • ab 9110 at a 1/200 dilution staining the mouse olineu cell line (oligodendrocyte precursor cell) by immunocytochemistry. The antibody was incubated with the cells for 30 minutes and then detected using a Cy5 conjugated goat anti-rabbit antibody.

    This image is courtesy of an Abreview submitted by Katarina Trajkovic on 15 March 2006

    See Abreview

References

This product has been referenced in:
  • He Y  et al. A Pap1-Oxs1 signaling pathway for disulfide stress in Schizosaccharomyces pombe. Nucleic Acids Res 45:106-114 (2017). IP . Read more (PubMed: 27664222) »
  • Jung J  et al. Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator. Elife 6:N/A (2017). WB, ICC/IF . Read more (PubMed: 28195531) »

See all 398 Publications for this product

Customer reviews and Q&As

My colleagues confirmed the antibody is non reactive with KLH and non transfected cells.

Although we have not tested this antibody specifically against proteins expressed in S. cerevisiae, it should recognise the protein in this species.

I have to confirm that we did not test ab9110 with multiple HA tag repeats in CHIP but only with a single tag.

But I would expect ab9110 to work also with multiple HA tag repeats.

Multiple freeze-thaw cycles may reduce the activity of this antibody. If you are concerned about losing activity by freezing this antibody, it can be stored in 50% glycerol at -20C. This will keep the antibody from freezing while allowing it to be stor...

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I can confirm that this primary antibody (ab9110) can be used for ELISA. Since it is not labelled it is important to use a compatible secondary antibody for detection which is conjugated to either to an enzyme or to a fluorochrome.

Yes, ab9110 is the unconjugated form of ab26228 (biotinylated form).

Both antibodies have not yet been tested in any IHC application (neither IHC-P or IHC-Fr).

Yes, you are correct, this antiserum was validated using our unique system of HA-tagging BORIS and injecting into Xenopus oocytes. The antiserum has not yet been tested by ChIP in human cells. However, this antiserum was tested by ChIP using the stan...

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Under the IF suggested dilution range for this antibody, the 1:40,000 dilution was a typo - it was supposed to be 1:4000. We have received some nice feedback from customers who have used this antibody in IF, and several have mentioned obtaining good re...

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This product will recognize the C-terminus, N-terminus and middle.

1-10 of 12 Q&A

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

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